LOCUS       pLenti-RAV             10991 bp    DNA     circular SYN 14-NOV-2024
DEFINITION  synthetic circular DNA
ACCESSION   VB240214-1123kqt
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10991)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 19, 2025 from SnapGene 8.0.2
            https://www.snapgene.com
COMMENT     Sequence Label: VB240214-1123kqt.
FEATURES             Location/Qualifiers
     source          1..10991
                     /organism="synthetic DNA construct"
     promoter        1..229
                     /label=RSV  promoter
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Strong promoter; drives 
                     transcriptionof viral RNA in packaging cells."
                     /note="/full_name=Rous sarcoma virus (RSV) 
                     enhancer/promoter"
                     /note="/name=RSV  promoter"
                     /note="/official_designation=RSV"
                     /note="/uuid=3f2ddca7f0cb4015b6abcec8a25c834e"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5' LTR-Delta-U3
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Allows transcription of viral RNA
                     andits packaging into virus."
                     /note="/full_name=HIV-1 truncated 5' LTR"
                     /note="/name=5' LTR-Delta-U3"
                     /note="/official_designation=Delta5' LTR"
                     /note="/uuid=44156a3083604321be44066133e8ecb9"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    521..565
                     /label=Psi
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Allows packaging of viral RNA 
                     intovirus."
                     /note="/full_name=HIV-1 psi packaging signal"
                     /note="/name=Psi"
                     /note="/official_designation=Psi"
                     /note="/uuid=284899413ff943a99f8210a11ad8c54e"
     misc_feature    1075..1308
                     /label=RRE
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Rev protein binding site that 
                     allowsRev-dependent nuclear export of viral RNA during 
                     viral packaging."
                     /note="/full_name=HIV-1 Rev response element"
                     /note="/name=RRE"
                     /note="/official_designation=RRE"
                     /note="/uuid=df07099e49674cf3aba3661b54e28bd7"
     primer_bind     1253..1271
                     /label=2647
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     1253..1271
                     /label=2648
                     /note="primer_bind	orchid	green	0	0"
     CDS             1493..1537
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     primer_bind     1591..1610
                     /label=2044
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     1661..1680
                     /label=2125
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     1671..1690
                     /label=2068
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(1680..1697)
                     /label=2440
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     1701..1720
                     /label=2057
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     1720..1740
                     /label=655
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     1721..1740
                     /label=2059
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(1780..1798)
                     /label=2066
                     /note="primer_bind	orchid	green	0	0"
     misc_feature    1803..1920
                     /label=cPPT
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Facilitates the nuclear import of
                     HIV-1cDNA through a central DNA flap."
                     /note="/full_name=Central polypurine tract"
                     /note="/name=cPPT"
                     /note="/official_designation=cPPT"
                     /note="/uuid=f7cb29985f9f41a7a5924d3462266f42"
     protein_bind    1938..1958
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_feature    1959..5652
                     /label={RAV}
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/name={RAV}"
                     /note="/official_designation={RAV}"
                     /note="/uuid=402822e70c9146829a2226340a4f2835"
     primer_bind     complement(1971..1990)
                     /label=2762
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(2074..2096)
                     /label=2992
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     2094..2113
                     /label=2771
                     /note="primer_bind	orchid	green	0	0"
     misc_feature    2592..3283
                     /label=Nr1d1 Ex1
     primer_bind     complement(2711..2727)
                     /label=741
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     2789..2805
                     /label=742
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     2794..2813
                     /label=2975
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(2871..2890)
                     /label=2976
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     2991..3010
                     /label=2985
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(3073..3092)
                     /label=2986
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     3073..3092
                     /label=2987
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     3107..3126
                     /label=2981
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     3150..3169
                     /label=2977
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(3182..3201)
                     /label=2988
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     3215..3234
                     /label=2983
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     3241..3260
                     /label=2979
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     3248..3267
                     /label=2972
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     3250..3270
                     /label=898
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     3250..3270
                     /label=958
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(3277..3303)
                     /label=2982
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(3280..3304)
                     /label=2984
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(3354..3373)
                     /label=2974
                     /note="primer_bind	orchid	green	0	0"
     misc_feature    4334..4467
                     /label=Nr1d1 Ex2 trunc
     primer_bind     complement(4349..4368)
                     /label=2978
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(4402..4421)
                     /label=1707
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(4430..4449)
                     /label=2973
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(4431..4450)
                     /label=2980
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     4467..4488
                     /label=2597
                     /note="primer_bind	orchid	green	0	0"
     misc_feature    4468..4473
                     /label=MluI
                     /note="/__SeqFeature__=True"
                     /note="/__level__=1"
                     /note="/direction=NONE"
                     /note="/name=MluI"
                     /note="/official_designation=MluI"
                     /note="/uuid=157d822145f14df2a23981abab96a158"
     primer_bind     4472..4497
                     /label=2073
                     /note="primer_bind	orchid	green	0	0"
     CDS             join(4474..4476,4477..5187)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             4474..5186
                     /label=Venus
                     /note="/Application Notes=Fast maturation and high 
                     tolerance toacidosis and Cl- compared to its predecessor 
                     YFP."
                     /note="/Brightness=156"
                     /note="/Full Name=Venus"
                     /note="/Maximum Emission (nm)=528"
                     /note="/Maximum Excitation (nm)=515"
                     /note="/References=Nat biotechnol. 20:87 (2002)"
                     /note="/Structure=Monomer (may form weak dimer)"
                     /note="/Subtype=Fluorescent reporter"
                     /note="/__SeqFeature__=True"
                     /note="/__level__=1"
                     /note="/description=Variant of yellow fluorescent protein 
                     (YFP)generated by mutagenesis; YFP is itself a variant of 
                     EGFP generated by mutagenesis"
                     /note="/name=Venus"
                     /note="/official_designation=Venus"
                     /note="/uuid=f8c7e087298f4a5984b69a6873f508a1"
     primer_bind     4474..4494
                     /label=594
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     4474..4494
                     /label=597
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     4474..4493
                     /label=927
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     4474..4490
                     /label=953
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     4474..4489
                     /label=2618
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(4479..4496)
                     /label=1927
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(4539..4559)
                     /label=2088
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     4702..4721
                     /label=734
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(4778..4797)
                     /label=735
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     4795..4817
                     /label=2608
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(4833..4852)
                     /label=1925
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(4845..4864)
                     /label=2624
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     4851..4870
                     /label=2625
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(4889..4906)
                     /label=2606
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     5009..5028
                     /label=2621
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5009..5028)
                     /label=2626
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5054..5073)
                     /label=1825
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     5092..5114
                     /label=1584
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     5097..5115
                     /label=729
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     5107..5126
                     /label=2627
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     5123..5142
                     /label=814
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     5123..5142
                     /label=838
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5123..5142)
                     /label=1023
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5141..5192)
                     /label=2471
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     5155..5177
                     /label=870
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5166..5186)
                     /label=596
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5166..5186)
                     /label=599
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5166..5186)
                     /label=595
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5166..5186)
                     /label=598
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5167..5186)
                     /label=2622
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5167..5186)
                     /label=1962
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5167..5186)
                     /label=2004
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5167..5186)
                     /label=1961
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5167..5186)
                     /label=2003
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5167..5182)
                     /label=2970
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5167..5182)
                     /label=2971
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5168..5186)
                     /label=1038
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5169..5192)
                     /label=2598
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5169..5186)
                     /label=1155
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5169..5186)
                     /label=1964
                     /note="primer_bind	orchid	green	0	0"
     misc_feature    5188..5193
                     /label=XbaI
                     /note="/__SeqFeature__=True"
                     /note="/__level__=1"
                     /note="/direction=NONE"
                     /note="/name=XbaI"
                     /note="/official_designation=XbaI"
                     /note="/uuid=fbb65844604642bf898f6d6bc0ba2215"
     misc_feature    5197..5217
                     /label=SV40-NLS
                     /label=nonstandard type:  Tag
                     /note="/Application Notes=Directs nuclear localization of 
                     taggedprotein."
                     /note="/Full Name=SV40 nuclear localization signal"
                     /note="/References=Cell. 39:499 (1984)"
                     /note="/Subtype=Signal peptide"
                     /note="/__SeqFeature__=True"
                     /note="/__level__=1"
                     /note="/description=Simian virus 40 nuclear localization 
                     signal"
                     /note="/name=SV40-NLS"
                     /note="/official_designation=SV40-NLS"
                     /note="/uuid=585a6ef53324445f8baca4b7b05cb10d"
     CDS             5197..5217
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5339..5356)
                     /label=928
                     /note="primer_bind	orchid	green	0	0"
     misc_feature    5359..5364
                     /label=XhoI
                     /note="/__SeqFeature__=True"
                     /note="/__level__=1"
                     /note="/direction=NONE"
                     /note="/name=XhoI"
                     /note="/official_designation=XhoI"
                     /note="/uuid=a8ebc898e58a420897d3b88d510c8102"
     misc_feature    5365..5652
                     /label=mNr1d1 3'UTR
                     /note="/__SeqFeature__=True"
                     /note="/__level__=1"
                     /note="/name=mNr1d1 3'UTR"
                     /note="/official_designation=mNr1d1 3'UTR"
                     /note="/uuid=a0d214e3478544689f2633dd97c82ca9"
     primer_bind     5395..5416
                     /label=1353
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     5651..5675
                     /label=574
                     /note="primer_bind	orchid	green	0	0"
     protein_bind    5652..5676
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     5652..5670
                     /label=1252
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     5653..5673
                     /label=1189
                     /note="primer_bind	orchid	green	0	0"
     promoter        5677..6187
                     /label=mPGK
                     /note="/Application Notes=Medium-strength promoter."
                     /note="/Full Name=Mouse phosphoglycerate kinase 1 promoter"
                     /note="/References=Gene. 60:65 (1987); PLoS One. 5:e10611 
                     (2010)"
                     /note="/Subtype=Mammalian ubiquitous promoter"
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Medium-strength promoter."
                     /note="/description=Mouse phosphoglycerate kinase 1 
                     promoter"
                     /note="/full_name=Mouse phosphoglycerate kinase 1 promoter"
                     /note="/name=mPGK"
                     /note="/official_designation=mPGK promoter"
                     /note="/uuid=bce93795b5fd4173b8b9abc832edad80"
     promoter        5684..6183
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5754..5773)
                     /label=1093
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(5966..5989)
                     /label=1482
                     /note="primer_bind	orchid	green	0	0"
     misc_feature    6188..6193
                     /label=kozak
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Facilitates translation 
                     initiation ofATG start codon downstream of the Kozak 
                     sequence."
                     /note="/full_name=Kozak"
                     /note="/name=kozak"
                     /note="/official_designation=Kozak"
                     /note="/uuid=5f18b25a1234443494a618363f09f052"
     CDS             6194..6793
                     /label=Puro
                     /note="/Application Notes=Allows cells to be resistant 
                     topuromycin."
                     /note="/Full Name=Puromycin resistance gene"
                     /note="/References=Gene. 62:121 (1988)"
                     /note="/Selection Drug=Puromycin"
                     /note="/Subtype=Drug selection marker"
                     /note="/Usage=Positive selection"
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Allows cells to be resistant 
                     topuromycin."
                     /note="/description=Puromycin resistance gene"
                     /note="/full_name=Puromycin resistance gene"
                     /note="/name=Puro"
                     /note="/official_designation=Puro"
                     /note="/uuid=39d7413208494d9291d10b21b13ca44c"
     primer_bind     complement(6197..6214)
                     /label=1842
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(6771..6790)
                     /label=914
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(6773..6793)
                     /label=926
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(6775..6793)
                     /label=1689
                     /note="primer_bind	orchid	green	0	0"
     protein_bind    complement(6794..6818)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(6796..6818)
                     /label=957
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(6801..6824)
                     /label=1253
                     /note="primer_bind	orchid	green	0	0"
     misc_feature    6832..7429
                     /label=WPRE
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Enhances virus stability in 
                     packagingcells, leading to higher titer of packaged virus; 
                     enhances higher expression of transgenes."
                     /note="/full_name=Woodchuck hepatitis virus 
                     posttranscriptionalregulatory element"
                     /note="/name=WPRE"
                     /note="/official_designation=WPRE"
                     /note="/uuid=0aedff8308474b369c622433538d7836"
     misc_feature    6836..7424
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(7307..7318)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7511..7745
                     /label=3' LTR-Delta-U3
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Allows packaging of viral RNA 
                     intovirus; self-inactivates the 5' LTR by a copying 
                     mechanism during viral genome integration; contains 
                     polyadenylation signal for transcription termination."
                     /note="/full_name=HIV-1 truncated 3' LTR"
                     /note="/name=3' LTR-Delta-U3"
                     /note="/official_designation=DeltaU3/3' LTR"
                     /note="/uuid=c958755414c34372b909e20e81ebcef6"
     LTR             7512..7745
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7817..7938
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    7818..7952
                     /label=SV40 early pA
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Allows transcription termination 
                     andpolyadenylation of mRNA transcribed by Pol II RNA 
                     polymerase."
                     /note="/full_name=SV40 early polyadenation signal"
                     /note="/name=SV40 early pA"
                     /note="/official_designation=SV40 early pA"
                     /note="/uuid=2fcb13a62e704d8095b188d9625a6a94"
     primer_bind     complement(7905..7925)
                     /label=792
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(7908..7938)
                     /label=2990
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(7931..7955)
                     /label=2442
                     /note="primer_bind	orchid	green	0	0"
     rep_origin      7978..8113
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8069..8088)
                     /label=2931
                     /note="primer_bind	orchid	green	0	0"
     promoter        complement(8134..8152)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8134..8152)
                     /label=349
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(8162..8178)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8163..8178)
                     /label=42
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(8182..8205)
                     /label=1058
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     complement(8303..8323)
                     /label=2435
                     /note="primer_bind	orchid	green	0	0"
     rep_origin      8320..8775
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     8501..8519
                     /label=700
                     /note="primer_bind	orchid	green	0	0"
     promoter        8801..8905
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8906..9766
                     /label=Ampicillin
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Allows E. coli to be resistant 
                     toampicillin."
                     /note="/full_name=Ampicillin resistance gene"
                     /note="/name=Ampicillin"
                     /note="/official_designation=Ampicillin"
                     /note="/uuid=6a7994dc58024d80b36e4a85ad2e1d99"
     rep_origin      9937..10525
                     /direction=RIGHT
                     /label=pUC ori
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Facilitates plasmid replication 
                     in E.coli; regulates high-copy plasmid number (500-700)."
                     /note="/full_name=pUC origin of replication"
                     /note="/name=pUC ori"
                     /note="/official_designation=pUC ori"
                     /note="/uuid=64a8cb3def9f49deb6752795c2d893a8"
     primer_bind     10641..10659
                     /label=2437
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     10710..10733
                     /label=2436
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     10735..10754
                     /label=2438
                     /note="primer_bind	orchid	green	0	0"
     protein_bind    10813..10834
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10849..10854,10855..10872,10873..10879)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     10880..10899
                     /label=1069
                     /note="primer_bind	orchid	green	0	0"
     protein_bind    10887..10903
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     10911..10927
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10911..10927
                     /label=43
                     /note="primer_bind	orchid	green	0	0"
     primer_bind     10946..10966
                     /label=891
                     /note="primer_bind	orchid	green	0	0"
     promoter        10948..10966
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
BASE COUNT         2587 a         3005 c         2831 g         2568 t
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgcta
     1801 gcttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
     1861 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt
     1921 actagtgatt atcggatcaa ctttgtatag aaaagttgcc ctctctctat gaggacctca
     1981 gagcactttc cctttgttct cctttttatt tcccagacaa agggaaatga ctcaccccaa
     2041 agtcacttga gtgggcagtg tggtgtcata taagcaaaca gggagtccct agggatatcc
     2101 ccgtgctatg ccagtcctac ccgtgggggc tggggttagg gtgaagaaga gaaaggggtt
     2161 ttatttacac tgcttgtcgc ccaccttgtc accatagcaa aagatgctta agttcttgat
     2221 ttaagggaaa tctctcctcc cagcttgtct ttctccccct agtcaccact aacctcaggg
     2281 tgaggtcagt tctccaatca cgtgaagctc tcacgtttgc taggtttgca gaaagggcct
     2341 tttagctttg atcttccacg gagcagtaaa gcttgccagg ccctccccag gaattcacat
     2401 gcccttgcca tacaaggctt tccaaacacg ccaccctgac tcttcagaaa accccacccc
     2461 agtccacttt cagctcctcc ccgctgccag caagcacttt tgcaaagcag aaaaggaata
     2521 ggcacactcc acctacattg tcagtagact acaaatccca acaatcctgg cggtgcgcag
     2581 gcgcacagat ctcaacgtgc cggctgctgg aaaagtgtgt cactggggca cgaggcgctc
     2641 cctggaatca catggtacct gctccagtgc cgcgtgcggc ccgggaaccc tgggctgctg
     2701 gcgcctgcgc agagccctct gtcccaggga aaggctcggg caaaaggcgg ttgagattgg
     2761 cagagtgaaa tattactgct gaggaaacgt agcagggcac acgtctctct ttgcgtcctg
     2821 gtgcctcatt tctccatcac ctacttactt cctggttgca gcttctcttc ctttgggact
     2881 tttgcacccg gagctccaga ttcattaccc tgcttcactg cggagccttc aggagtggca
     2941 tccggtgcac tgcagagacc cgactctcct tgctacctta tagccagaac tgctgcaggc
     3001 tgattcttca cacacacact ctctgctctt cccatgcaaa tcagatctca ggtgcctcag
     3061 cgtcctaccc ttctggaggg ctgcagtata gccaccccaa gaccttactg ctcagtgcct
     3121 ggaatcctga ttgcgaactg cggggctcac tcgtctccct cagccattgc ccagggggcg
     3181 agagaggcca tcacaacctc cagtttgtgt caaggtccag tttgaatgac cgctttcagc
     3241 tggtgaagac atgacgaccc tggactccaa taacaacaca ggtactgaga ttcttatctt
     3301 tgctcagggc tcacatggct gcagagccgg gcccacgtgc tgcatttgtt ttcattcagt
     3361 agaagccgtg gccgccatct cattggtcat tgcctcccgg cctcgttaca taatgagctc
     3421 cgcccctcaa gttgccggag caacgcttgt cgccttctct cctcccctca gttgtcccgg
     3481 cccagcttgg ttttagggcc agcctacgac ccaggacgcc cgggaggggc ggggcgaggg
     3541 cggggttttc cttgcgacag aagggaagcg gagaaggcgg tgcttcgagt tccggggttc
     3601 ctcgttgggg tgccacgtgc gaggggcaca cgtggagtgg ggacgtgagg ccagctgagg
     3661 gccaggctcc cgccccgccc ttctccgcct agatctgggt gaagctgggc tgactcaccg
     3721 cccccgcctg gcttcttttc ctctcagaaa ataacccatc ctccaaccca gccgtgaaac
     3781 catagccggg taaccattct gtgacttatt ttacaggcag ggttcaccct gaccaacctt
     3841 gagctatttt aaggttgccc tgcctggttt agtgtgtgtc agtgggtgag caagacatgt
     3901 gcttctgtcg actcggtggc atggcatgtg tcacctcaca cacgtgtctg aggatcctcc
     3961 cctttcccca ctccacccat ggccggccag gctaagtact tggcagaggt gtggcagaca
     4021 agtggccctt ctcttctgtg cataccacag gagccgaatc acctggcccg caatgtaacc
     4081 agagggacag agccgctggc tgagtggcta tagagctaaa gttaccaccg ggctgagaag
     4141 gaaggggaat tcctaaatcc ctagaatgga tactttgttt ctccctttca tctcctgcaa
     4201 ctcatctgag tcttatctgt cacatttcta agtctcttgt gactcatctc tcagaggcca
     4261 gagccaggct gaacagagag tccttccctg cagtactcta tgagaatatg tctttgggtg
     4321 ttctcttatt taggtggtgt tatcacctac attggctcta gtggctcctc cccgagccgg
     4381 accagcccgg agtccctcta cagtgacagc tccaatggca gcttccagtc cctgactcaa
     4441 ggttgtccca catacttccc accatcaacg cgtgtgagca agggcgagga gctgttcacc
     4501 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     4561 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc
     4621 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcctgcag
     4681 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     4741 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     4801 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     4861 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     4921 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     4981 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     5041 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccgc cctgagcaaa
     5101 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     5161 actctcggca tggacgagct gtacaaatct agacctccaa aaaagaagag aaaggtagaa
     5221 gaccccttgt acaagagcca tggcttcccg ccggctgtgg ctgctcagga tgatggcacg
     5281 ctgcccatgt cctgtgccca ggagagcggg atggaccgtc accctgcagc ctgtgctcct
     5341 gctaggatca atgtgtgact cgagcccgcc cggccggcct tctgccgctg cccccttgta
     5401 cagaatcgaa ctctgcactt ctctctcctt tacgagacga aaaggaaaag caaaccagaa
     5461 tcttatttat attgttataa aatattccaa gatgagcctc tggccccctg agccttcttg
     5521 taaataactc ttcccccacc cccaccgcca tgctcccatc ctcccctatt taaaccactc
     5581 ttgctctctc caccctcctc tggcccctcg atttgttctg ttcctgtctc aaatccaata
     5641 gttcacagct gacaagtttg tacaaaaaag caggctttct accgggtagg ggaggcgctt
     5701 ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca
     5761 agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc
     5821 tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc
     5881 ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg
     5941 cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag
     6001 cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg
     6061 ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc
     6121 tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt
     6181 tcgacctgcc accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     6241 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     6301 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     6361 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     6421 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     6481 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     6541 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     6601 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     6661 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     6721 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     6781 gcccggtgcc tgaacccagc tttcttgtac aaagtggtga taatcgaatt ccgataatca
     6841 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     6901 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     6961 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     7021 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     7081 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     7141 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     7201 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     7261 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     7321 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     7381 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcggg aattcccgcg
     7441 gttcgcttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     7501 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     7561 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     7621 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     7681 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     7741 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     7801 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     7861 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     7921 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
     7981 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     8041 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     8101 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
     8161 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     8221 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     8281 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     8341 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     8401 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     8461 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     8521 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     8581 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     8641 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     8701 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     8761 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     8821 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     8881 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     8941 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     9001 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     9061 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     9121 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     9181 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     9241 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     9301 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     9361 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     9421 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     9481 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     9541 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     9601 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     9661 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     9721 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     9781 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     9841 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     9901 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     9961 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
    10021 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
    10081 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
    10141 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
    10201 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
    10261 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
    10321 gcgtgagcta tgagaaagcg ccacgcttcc cgaagagaga aaggcggaca ggtatccggt
    10381 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
    10441 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
    10501 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
    10561 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
    10621 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
    10681 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
    10741 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
    10801 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
    10861 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
    10921 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga
    10981 gctgcaagct t
//
