LOCUS       Exported                5159 bp ds-DNA     circular SYN 01-SEP-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5159)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Mar 25, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Annotated with pLannotate v1.1.0
FEATURES             Location/Qualifiers
     source          1..5159
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    3..132
                     /label=AAV2 ITR (alternate)
     repeat_region   14..41
                     /label=rAAV2 ITR (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=19.9"
                     /note="/other=repeat_region"
     repeat_region   70..97
                     /label=rAAV2 ITR (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=19.9"
                     /note="/other=repeat_region"
     promoter        166..613
                     /label=rhSyn promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     primer_bind     591..610
                     /label=hSyn-F
     misc_feature    626..659
                     /label=FRT5
     misc_feature    626..659
                     /label=FRT5
     misc_feature    722..769
                     /label=rFRT
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     misc_feature    782..815
                     /label=rloxP
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     misc_feature    complement(816..907)
                     /label=T3H48 ribozyme
     ncRNA           complement(817..866)
                     /label=rHammerhead 1
                     /note="pLannotate"
                     /note="/database=Rfam"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=108.7"
                     /note="/other=ncRNA"
     misc_feature    908..941
                     /label=rloxP
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     primer_bind     complement(932..956)
                     /label=EGFP-C
     CDS             complement(956..1675)
                     /codon_start=1
                     /label=rEGFP
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(959..1675)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     971..990
                     /label=eGFP_3
     primer_bind     974..992
                     /label=AH-Split-ATG-1209-F
     primer_bind     complement(974..992)
                     /label=AH-split-ATG-1209-R
     primer_bind     complement(1001..1022)
                     /label=EGFPCnew
     primer_bind     1033..1052
                     /label=eGFP_2
     primer_bind     1065..1084
                     /label=eGFPF1
     primer_bind     complement(1188..1206)
                     /label=eGFPR1
     primer_bind     complement(1435..1454)
                     /label=eGFP_4
     primer_bind     1435..1454
                     /label=eGFP_4R
     primer_bind     complement(1536..1555)
                     /label=eGFP_1
     regulatory      1672..1681
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    complement(1700..1733)
                     /label=FRT5
     misc_feature    complement(1700..1733)
                     /label=FRT5
     protein_bind    complement(1796..1843)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     protein_bind    complement(1796..1829)
                     /label=rFRT (minimal)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     primer_bind     complement(1884..1905)
                     /label=WPRE-5
     misc_feature    1894..2051
                     /label=rWPRE (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=96.2"
                     /note="/match_length=39.1"
                     /note="/other=misc_feature"
     misc_feature    1895..2137
                     /label=wPRE3 (estimated)
     primer_bind     complement(1947..1970)
                     /label=WPRE_2
     primer_bind     complement(1947..1967)
                     /label=WPRE_Rev
     misc_feature    2057..2141
                     /label=rWPRE (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=14.4"
                     /note="/other=misc_feature"
     polyA_signal    complement(2140..2183)
                     /label=rSV40 poly(A) signal (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=53.7"
                     /note="/other=polyA_signal"
     misc_feature    2140..2183
                     /label=SV40 late polyA
     misc_feature    2186..2320
                     /label=SL (estimated)
     polyA_signal    complement(2187..2254)
                     /label=rSV40 poly(A) signal (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=82.9"
                     /note="/other=polyA_signal"
     misc_feature    2189..2232
                     /label=SV40 late polyA
     repeat_region   2340..2480
                     /label=rAAV2 ITR
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=repeat_region"
     repeat_region   2340..2469
                     /label=AAV2 ITR
     primer_bind     complement(2520..2536)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2544..2560)
                     /label=rlac operator
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     promoter        complement(2568..2598)
                     /label=rlac promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     protein_bind    complement(2613..2634)
                     /label=rCAP binding site
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     CDS             complement(2647..2739)
                     /codon_start=1
                     /label=rlacI (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=8.6"
                     /note="/other=CDS"
                     /translation="APNTQTASPRALADSLMQLARQVSRLESGQ"
     rep_origin      complement(2922..3510)
                     /direction=LEFT
                     /label=rori
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.8"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
     CDS             complement(3681..4541)
                     /codon_start=1
                     /label=rAmpR
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.7"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4542..4646)
                     /label=rAmpR promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     CDS             4741..4776
                     /codon_start=1
                     /label=rrop (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=18.8"
                     /note="/other=CDS"
                     /translation="LARFGDDGENL"
     misc_feature    4878..4977
                     /label=rbom (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=70.9"
                     /note="/other=misc_feature"
     CDS             complement(4983..5140)
                     /codon_start=1
                     /label=rlacZ-alpha (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=90.8"
                     /note="/other=CDS"
                     /translation="IHWPSFYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPN
                     SCAA*MA"
     primer_bind     5120..5136
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 atcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct
       61 ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca
      121 ctaggggttc ctgcggccgc agatgctgca cgggcccgcg atcgcagtgc aagtgggttt
      181 taggaccagg atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga
      241 caagcaccca acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa
      301 caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc
      361 cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc
      421 cggtcccccg caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc
      481 agccggaccg caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg
      541 cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg
      601 cctgagagcg cagggtaccg ctagcgaagt tcctattctt caaaaggtat aggaacttca
      661 tcaaaatagg aagaccaatg cttcaccatc gacccgaatt gccaagcatc accatcgaca
      721 cgaagttcct attccgaagt tcctattctc tagaaagtat aggaacttcg tcgaccttaa
      781 gataacttcg tatagcatac attatacgaa gttataaaaa caaaaagaaa agcgcgtttc
      841 gtcctatttg ggactcgtca gctggatgta cgcgtttccg cgaatccagg acgcgcaaag
      901 aaagaaaata acttcgtata gcatacatta tacgaagtta tgaattcatg catgcttact
      961 tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
     1021 tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctgggtgctc aggtagtggt
     1081 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
     1141 gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct
     1201 tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc
     1261 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
     1321 tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga
     1381 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
     1441 tgtggtcggg gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg
     1501 gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg
     1561 tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca
     1621 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg
     1681 cactagtacc ggtctcgagg aagttcctat accttttgaa gaataggaac ttcatcaaaa
     1741 taggaagacc aatgcttcac catcgacccg aattgccaag catcaccatc gacacgaagt
     1801 tcctatactt tctagagaat aggaacttcg gaatagtaac ttccgtacga agctttctag
     1861 agcttagccg attcgatatt cgatgcccga gataatcaac ctctggatta caaaatttgt
     1921 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     1981 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2041 aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     2101 acaggggctc ggctgttggg cactgacaat tccgtggtgt ttatttgtga aatttgtgat
     2161 gctattgctt tatttgtaac catctagctt tatttgtgaa atttgtgatg ctattgcttt
     2221 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2281 gtttcaggtt cagggggaga tgtgggaggt tttttaaagc ggccggtaac cggagctcca
     2341 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     2401 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     2461 agcgcgcagc tgcctgcagg attggatccc aatggcgcgc cgagcttggc gtaatcatgg
     2521 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     2581 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2641 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2701 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2761 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2821 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2881 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2941 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3001 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3061 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3121 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3181 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3241 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3301 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3361 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3421 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3481 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3541 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3601 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3661 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3721 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3781 gagggcttac catctggccc cagtgctgca atgataccgc gagaaccacg ctcaccggct
     3841 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3901 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3961 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4021 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4081 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4141 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4201 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4261 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4321 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4381 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4441 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4501 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4561 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4621 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     4681 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gcccttttgt
     4741 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagactgtc
     4801 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     4861 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     4921 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     4981 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     5041 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     5101 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgacgcgtat tgggatcca
//
