LOCUS       Exported                8179 bp ds-DNA     circular SYN 29-JAN-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 8179)
  AUTHORS   Sho Yoshimatsu
  TITLE     Direct Submission
  JOURNAL   Exported Sep 21, 2020 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8179
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     primer_bind     1..20
                     /label=pCRstart-5
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     107..128
                     /label=CAP-5
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     149..168
                     /label=Lac pro-5
     primer_bind     181..200
                     /label=Lac ope-5
     protein_bind    181..197
                     /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     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             217..336
                     /codon_start=1
                     /gene="lacZ (fragment)"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASSLVPSSDPLVTAASVLEFAL"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     239..257
                     /label=SP6-5
     gene            337..1894
                     /gene="DDX4"
                     /label=DDX4
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon."
                     /db_xref="GeneID:100390219"
     mRNA            join(337..526,666..776)
                     /gene="DDX4"
                     /product="DEAD-box helicase 4, transcript variant X2"
                     /label=DDX4 mRNA
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon. Supporting evidence 
                     includes similarity to: 92 ESTs, 1 Protein, and 100% 
                     coverage of the annotated genomic feature by RNAseq 
                     alignments, including 5 samples with support for all 
                     annotated introns"
                     /db_xref="GeneID:100390219"
     mRNA            join(337..526,666..776)
                     /gene="DDX4"
                     /product="DEAD-box helicase 4, transcript variant X1"
                     /label=DDX4 mRNA
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon. Supporting evidence 
                     includes similarity to: 89 ESTs, and 100% coverage of the 
                     annotated genomic feature by RNAseq alignments, including 2
                     samples with support for all annotated introns"
                     /db_xref="GeneID:100390219"
     mRNA            join(337..526,666..776)
                     /gene="DDX4"
                     /product="DEAD-box helicase 4, transcript variant X4"
                     /label=DDX4 mRNA
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon. Supporting evidence 
                     includes similarity to: 32 ESTs, 1 Protein, and 100% 
                     coverage of the annotated genomic feature by RNAseq 
                     alignments, including 5 samples with support for all 
                     annotated introns"
                     /db_xref="GeneID:100390219"
     mRNA            join(337..526,666..776)
                     /gene="DDX4"
                     /product="DEAD-box helicase 4, transcript variant X3"
                     /label=DDX4 mRNA
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon. Supporting evidence 
                     includes similarity to: 29 ESTs, and 100% coverage of the 
                     annotated genomic feature by RNAseq alignments, including 2
                     samples with support for all annotated introns"
                     /db_xref="GeneID:100390219"
     CDS             join(337..526,666..776)
                     /codon_start=1
                     /gene="DDX4"
                     /product="probable ATP-dependent RNA helicase DDX4 isoform 
                     X4"
                     /label=DDX4
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon."
                     /db_xref="GeneID:100390219"
                     /protein_id="XP_008990301.1"
                     /translation="AARGLDIENVQHVINFDLPSTIDEYVHRIGRTGRCGNTGRAISFF
                     DPESDNHLAQPLIKVLTDAQQDVPAWLEEIAFSAYIPGFSGSTRGSVFASVDTRK"
     CDS             join(337..526,666..776)
                     /codon_start=1
                     /gene="DDX4"
                     /product="probable ATP-dependent RNA helicase DDX4 isoform 
                     X3"
                     /label=DDX4
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon."
                     /db_xref="GeneID:100390219"
                     /protein_id="XP_017825400.1"
                     /translation="AARGLDIENVQHVINFDLPSTIDEYVHRIGRTGRCGNTGRAISFF
                     DPESDNHLAQPLIKVLTDAQQDVPAWLEEIAFSAYIPGFSGSTRGSVFASVDTRK"
     CDS             join(337..526,666..776)
                     /codon_start=1
                     /gene="DDX4"
                     /product="probable ATP-dependent RNA helicase DDX4 isoform 
                     X2"
                     /label=DDX4
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon."
                     /db_xref="GeneID:100390219"
                     /protein_id="XP_008990298.1"
                     /translation="AARGLDIENVQHVINFDLPSTIDEYVHRIGRTGRCGNTGRAISFF
                     DPESDNHLAQPLIKVLTDAQQDVPAWLEEIAFSAYIPGFSGSTRGSVFASVDTRK"
     CDS             join(337..526,666..776)
                     /codon_start=1
                     /gene="DDX4"
                     /product="probable ATP-dependent RNA helicase DDX4 isoform 
                     X1"
                     /label=DDX4
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon."
                     /db_xref="GeneID:100390219"
                     /protein_id="XP_017825399.1"
                     /translation="AARGLDIENVQHVINFDLPSTIDEYVHRIGRTGRCGNTGRAISFF
                     DPESDNHLAQPLIKVLTDAQQDVPAWLEEIAFSAYIPGFSGSTRGSVFASVDTRK"
     primer_bind     337..359
                     /label=cjDDX4gg2-F3
     primer_bind     686..708
                     /label=cjDDX4gg2-F2
     primer_bind     859..881
                     /label=cjDDX4gg-F2
     misc_feature    895..1894
                     /label=cjDDX4 5'arm 1k
     primer_bind     1529..1544
                     /label=DDX4-fus-F1
     CDS             1811..1894
                     /codon_start=1
                     /gene="DDX4"
                     /product="probable ATP-dependent RNA helicase DDX4 isoform 
                     X1"
                     /label=DDX4
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon."
                     /db_xref="GeneID:100390219"
                     /protein_id="XP_017825399.1"
                     /translation="NYQGKSTLNTAGFSSSQMSNPVDDESWD"
     CDS             1811..1894
                     /codon_start=1
                     /gene="DDX4"
                     /product="probable ATP-dependent RNA helicase DDX4 isoform 
                     X2"
                     /label=DDX4
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon."
                     /db_xref="GeneID:100390219"
                     /protein_id="XP_008990298.1"
                     /translation="NYQGKSTLNTAGFSSSQMSNPVDDESWD"
     CDS             1811..1894
                     /codon_start=1
                     /gene="DDX4"
                     /product="probable ATP-dependent RNA helicase DDX4 isoform 
                     X3"
                     /label=DDX4
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon."
                     /db_xref="GeneID:100390219"
                     /protein_id="XP_017825400.1"
                     /translation="NYQGKSTLNTAGFSSSQMSNPVDDESWD"
     CDS             1811..1894
                     /codon_start=1
                     /gene="DDX4"
                     /product="probable ATP-dependent RNA helicase DDX4 isoform 
                     X4"
                     /label=DDX4
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon."
                     /db_xref="GeneID:100390219"
                     /protein_id="XP_008990301.1"
                     /translation="NYQGKSTLNTAGFSSSQMSNPVDDESWD"
     mRNA            1811..1894
                     /gene="DDX4"
                     /product="DEAD-box helicase 4, transcript variant X3"
                     /label=DDX4 mRNA
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon. Supporting evidence 
                     includes similarity to: 29 ESTs, and 100% coverage of the 
                     annotated genomic feature by RNAseq alignments, including 2
                     samples with support for all annotated introns"
                     /db_xref="GeneID:100390219"
     mRNA            1811..1894
                     /gene="DDX4"
                     /product="DEAD-box helicase 4, transcript variant X4"
                     /label=DDX4 mRNA
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon. Supporting evidence 
                     includes similarity to: 32 ESTs, 1 Protein, and 100% 
                     coverage of the annotated genomic feature by RNAseq 
                     alignments, including 5 samples with support for all 
                     annotated introns"
                     /db_xref="GeneID:100390219"
     mRNA            1811..1894
                     /gene="DDX4"
                     /product="DEAD-box helicase 4, transcript variant X1"
                     /label=DDX4 mRNA
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon. Supporting evidence 
                     includes similarity to: 89 ESTs, and 100% coverage of the 
                     annotated genomic feature by RNAseq alignments, including 2
                     samples with support for all annotated introns"
                     /db_xref="GeneID:100390219"
     mRNA            1811..1894
                     /gene="DDX4"
                     /product="DEAD-box helicase 4, transcript variant X2"
                     /label=DDX4 mRNA
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon. Supporting evidence 
                     includes similarity to: 92 ESTs, 1 Protein, and 100% 
                     coverage of the annotated genomic feature by RNAseq 
                     alignments, including 5 samples with support for all 
                     annotated introns"
                     /db_xref="GeneID:100390219"
     misc_feature    1837..1875
                     /label=cjDDX4term 5'microhomology
     primer_bind     complement(1863..1900)
                     /label=DDX4-fus-R1(3)
     primer_bind     complement(1863..1900)
                     /label=DDX4-fus-R1(2)
     primer_bind     complement(1863..1892)
                     /label=DDX4-fus-R1
     misc_feature    1877..1894
                     /label=cjDDX4term 3'microhomology
     primer_bind     1895..1920
                     /label=P2A-NheI-F
     CDS             1901..1957
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     1901..1919
                     /label=FokI-P2A-5
     primer_bind     1901..1918
                     /label=rTAM2-P2A-F
     primer_bind     complement(1918..1943)
                     /label=P2A-deltaEsp3I-3
     primer_bind     1929..1954
                     /label=P2A-deltaEsp3I-5
     primer_bind     1931..1971
                     /label=T2A-tdTomato-5
     primer_bind     complement(1934..1953)
                     /label=P2A-3
     primer_bind     1941..1978
                     /label=P2A-tomato-F
     primer_bind     1941..1975
                     /label=P2A-GFP-5
     CDS             1958..3382
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYE
                     GTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMN
                     FEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVL
                     KGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYER
                     SEGRHHLFLYGMDELYK"
     primer_bind     1958..1972
                     /label=EcoRI-NotI-NheI-BamHI-k-EGFP-F
     misc_feature    1959..1960
                     /label=ttaa
     primer_bind     1961..1975
                     /label=FP-Fwd
     primer_bind     1961..1974
                     /label=NheI-k-Venus-F
     primer_bind     1961..1972
                     /label=BamHI-k-EGFP-F
     primer_bind     1961..1972
                     /label=Ac-EGFP-F
     primer_bind     1961..1970
                     /label=BamHI-mCerulean-F
     primer_bind     1961..1970
                     /label=pKObam-FP-F
     primer_bind     2098..2111
                     /label=BamHI-mCerulean-F
     primer_bind     2098..2111
                     /label=pKObam-FP-F
     primer_bind     2344..2354
                     /label=EcoRI-hH1foo-F
     primer_bind     complement(2353..2378)
                     /label=tdTomato-delBbsI-3
     primer_bind     2364..2389
                     /label=tdTomato-delBbsI-5
     primer_bind     2410..2420
                     /label=EcoRI-hH1foo-F
     primer_bind     2412..2424
                     /label=Puro-AatII-5
     primer_bind     2417..2440
                     /label=mCherry-Delta-S1-F
     primer_bind     complement(2512..2529)
                     /label=mCherry-Delta-S2-R
     primer_bind     complement(2651..2666)
                     /label=tdTlinker-R
     primer_bind     2651..2666
                     /label=tdTlinker-F
     primer_bind     2824..2837
                     /label=BamHI-mCerulean-F
     primer_bind     2824..2837
                     /label=pKObam-FP-F
     primer_bind     3070..3080
                     /label=EcoRI-hH1foo-F
     primer_bind     complement(3079..3104)
                     /label=tdTomato-delBbsI-3
     primer_bind     3090..3115
                     /label=tdTomato-delBbsI-5
     primer_bind     3136..3146
                     /label=EcoRI-hH1foo-F
     primer_bind     3138..3150
                     /label=Puro-AatII-5
     primer_bind     3143..3166
                     /label=mCherry-Delta-S1-F
     primer_bind     complement(3238..3255)
                     /label=mCherry-Delta-S2-R
     primer_bind     3356..3376
                     /label=FP-BsrGI-F
     primer_bind     complement(3361..3385)
                     /label=NotI-loxV-Venus-R
     primer_bind     complement(3363..3386)
                     /label=Ac-EGFP-R
     primer_bind     complement(3364..3391)
                     /label=bGHpA-SalI-FP-R
     primer_bind     complement(3364..3384)
                     /label=EGFP-NotI-R
     primer_bind     complement(3365..3391)
                     /label=pKOsal-FP-R
     primer_bind     complement(3365..3382)
                     /label=EGFP-end-3
     primer_bind     complement(3365..3382)
                     /label=tdTomato-3
     primer_bind     complement(3366..3391)
                     /label=SalI-mCerulean-R
     primer_bind     complement(3367..3382)
                     /label=EGFP-5'TR-3
     primer_bind     complement(3369..3382)
                     /label=mCherry-SacI-3
     primer_bind     complement(3370..3384)
                     /label=BglII-tdT-R
     primer_bind     3386..3417
                     /label=NheI-SalI-cjDDX4-3'arm-F
     misc_feature    3392..4391
                     /label=cjDDX4 3'arm 1k
     gene            3392..4004
                     /gene="DDX4"
                     /label=DDX4
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon."
                     /db_xref="GeneID:100390219"
     mRNA            3392..4004
                     /gene="DDX4"
                     /product="DEAD-box helicase 4, transcript variant X2"
                     /label=DDX4 mRNA
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon. Supporting evidence 
                     includes similarity to: 92 ESTs, 1 Protein, and 100% 
                     coverage of the annotated genomic feature by RNAseq 
                     alignments, including 5 samples with support for all 
                     annotated introns"
                     /db_xref="GeneID:100390219"
     mRNA            3392..4004
                     /gene="DDX4"
                     /product="DEAD-box helicase 4, transcript variant X1"
                     /label=DDX4 mRNA
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon. Supporting evidence 
                     includes similarity to: 89 ESTs, and 100% coverage of the 
                     annotated genomic feature by RNAseq alignments, including 2
                     samples with support for all annotated introns"
                     /db_xref="GeneID:100390219"
     mRNA            3392..4004
                     /gene="DDX4"
                     /product="DEAD-box helicase 4, transcript variant X4"
                     /label=DDX4 mRNA
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon. Supporting evidence 
                     includes similarity to: 32 ESTs, 1 Protein, and 100% 
                     coverage of the annotated genomic feature by RNAseq 
                     alignments, including 5 samples with support for all 
                     annotated introns"
                     /db_xref="GeneID:100390219"
     mRNA            3392..4004
                     /gene="DDX4"
                     /product="DEAD-box helicase 4, transcript variant X3"
                     /label=DDX4 mRNA
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon. Supporting evidence 
                     includes similarity to: 29 ESTs, and 100% coverage of the 
                     annotated genomic feature by RNAseq alignments, including 2
                     samples with support for all annotated introns"
                     /db_xref="GeneID:100390219"
     misc_feature    3392..3413
                     /label=cjDDX4term 3'microhomology
     CDS             3392..3394
                     /codon_start=1
                     /gene="DDX4"
                     /product="probable ATP-dependent RNA helicase DDX4 isoform 
                     X4"
                     /label=DDX4
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon."
                     /db_xref="GeneID:100390219"
                     /protein_id="XP_008990301.1"
                     /translation=""
     CDS             3392..3394
                     /codon_start=1
                     /gene="DDX4"
                     /product="probable ATP-dependent RNA helicase DDX4 isoform 
                     X3"
                     /label=DDX4
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon."
                     /db_xref="GeneID:100390219"
                     /protein_id="XP_017825400.1"
                     /translation=""
     CDS             3392..3394
                     /codon_start=1
                     /gene="DDX4"
                     /product="probable ATP-dependent RNA helicase DDX4 isoform 
                     X2"
                     /label=DDX4
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon."
                     /db_xref="GeneID:100390219"
                     /protein_id="XP_008990298.1"
                     /translation=""
     CDS             3392..3394
                     /codon_start=1
                     /gene="DDX4"
                     /product="probable ATP-dependent RNA helicase DDX4 isoform 
                     X1"
                     /label=DDX4
                     /note="Derived by automated computational analysis using 
                     gene prediction method: Gnomon."
                     /db_xref="GeneID:100390219"
                     /protein_id="XP_017825399.1"
                     /translation=""
     primer_bind     3406..3428
                     /label=Primer 3
     primer_bind     complement(3575..3600)
                     /label=HpaI-cjDDX4-3'arm-R2
     primer_bind     complement(3578..3620)
                     /label=DDX4-fus-R2
     primer_bind     complement(3578..3603)
                     /label=HpaI-cjDDX4-3'arm-R
     primer_bind     complement(3916..3938)
                     /label=cjDDX4gg-R1
     primer_bind     complement(4635..4654)
                     /label=cjDDX4gg-R2
     primer_bind     complement(4974..4996)
                     /label=cjDDX4gg2-R1
     CDS             4997..5236
                     /codon_start=1
                     /gene="lacZ (fragment)"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="KGEFCRYPSHWRPLEHASRGPNSPYSESYYNSLAVVLQRRDWENP
                     GVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSL"
     promoter        complement(5067..5085)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5067..5085)
                     /label=T7-5
     primer_bind     complement(5092..5108)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5137..5156)
                     /label=pCR-BEnd-3
     CDS             5246..5548
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(5273..5296)
                     /label=ccdB-3
     promoter        5759..5808
                     /label=NEO KAN promoter
     primer_bind     complement(5780..5799)
                     /label=NEOKANp-3
     CDS             5897..6691
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5897..5916)
                     /label=Neo-start-3
     primer_bind     6672..6691
                     /label=Neo-end-5
     promoter        6825..6853
                     /label=AmpR promoter
     primer_bind     6825..6844
                     /label=AmpP-5
     CDS             6898..7272
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     primer_bind     complement(7104..7122)
                     /label=NotI-loxV-Venus-R
     primer_bind     7104..7119
                     /label=FP-BsrGI-F
     primer_bind     complement(7109..7122)
                     /label=SalI-mCerulean-R
     primer_bind     complement(7110..7122)
                     /label=EGFP-5'TR-3
     rep_origin      7410..7998
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7666..7687
                     /label=Ori-5
     primer_bind     7979..7998
                     /label=Ori-end-5
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
      301 ctagtaacgg ccgccagtgt gctggaattc gcccttagct gccagagggc tagatattga
      361 aaatgtacag catgttatca attttgatct tccttctacc attgatgaat atgttcatcg
      421 aattgggcgt actggtcgtt gtgggaatac tggcagagca atttcctttt ttgatcctga
      481 atcggataac catttagcac agcctctaat aaaagtatta acagatgtaa gttaaacttt
      541 taaaacagaa tggatagttt tcctttgttg ttgaaagtag acattttatg tatatgtata
      601 taacaagtaa acttttccaa tctttaagac tttatgttgc atatgaagtc attttttttt
      661 ttaaggctca acaggatgtt cctgcatggc tggaagaaat tgcctttagt gcatacattc
      721 ctggcttcag tggtagtaca agaggaagtg tatttgcatc agttgatact agaaaggtta
      781 gtagaaagga agaattcaga atttggattt cagttactct ttgtaaacgt tatgcttaat
      841 attatgaagt aactataata atcagcccat tgttttcatg cctagaaggt agctacttta
      901 ttatttcatt aacattttat gtatatacac atttttggtt ttaaaggtca ggaattaact
      961 tttgagattt atacccaaac agtctccttt cccagatttt aaattttaat ttggaatctt
     1021 ctaaacttct aattgaatta tttaatttct gaattctgac aacattaata taaaatacag
     1081 gacctattag aattacaagt cttttctttt tttttttttt ttttttttga gatagtttgt
     1141 ggctctcttg tcgaggctgg agtgcagtgg catgatctca gctcgctgca acctctgcct
     1201 cccaggttca agcaattctt gtgcctcaag tgtatctggg attacaggca tgtaccacca
     1261 tcctcagctg tttttttttt tgtactttta atagagatag gatttcacca tcttggctag
     1321 gctggtcttg aactcctggc ctcaagtgat ccacctgcct cagcctccca aattactggg
     1381 attataggca tgagccactg tgcccagcca caaggctacg agtctttttt ggtatgtcag
     1441 gtttttatag gcagcaccac ttttatacag aaacagtaag catgctaatt aaatatttat
     1501 gcttattcat tgagtgtagt ccattaagtt ttccccactt gccaaaacaa ttggcttaaa
     1561 aaatattttt cttgagacat tttttgcaat ttagacttcc tggtttgaca tgagcctttt
     1621 tttcaagtga atcagattta gtgagaatga agttctttgt atatttgggg aagacatggt
     1681 agataccttg agtccttgct ctcattctga gtgacgtaac tttaaagttg tcataaaatt
     1741 aaatgtttgt ttcttttctt ttcttttttc tttctttgtc ttctttcttt cttttttttt
     1801 ttttttaaag aactaccagg gcaagagcac tttgaacaca gcaggatttt cttcttcaca
     1861 aatgtccaat cctgtggacg acgaatcatg ggatgctagc gccacgaact tctctctgtt
     1921 aaagcaagca ggagacgtgg aagaaaaccc cggtcctgtt agcaagggcg aggaggtcat
     1981 caaagagttc atgcgcttca aggtgcgcat ggagggctcc atgaacggcc acgagttcga
     2041 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     2101 gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt tcatgtacgg
     2161 ctccaaggcg tacgtgaagc accccgccga catccccgat tacaagaagc tgtccttccc
     2221 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggtctgg tgaccgtgac
     2281 ccaggactcc tccctgcagg acggcacgct gatctacaag gtgaagatgc gcggcaccaa
     2341 cttccccccc gacggccccg taatgcagaa aaagaccatg ggctgggagg cctccaccga
     2401 gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc caccaggccc tgaagctgaa
     2461 ggacggcggc cactacctgg tggagttcaa gaccatctac atggccaaga agcccgtgca
     2521 actgcccggc tactactacg tggacaccaa gctggacatc acctcccaca acgaggacta
     2581 caccatcgtg gaacagtacg agcgctccga gggccgccac cacctgttcc tggggcatgg
     2641 caccggcagc accggcagcg gcagctccgg caccgcctcc tccgaggaca acaacatggc
     2701 cgtcatcaaa gagttcatgc gcttcaaggt gcgcatggag ggctccatga acggccacga
     2761 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     2821 gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcat
     2881 gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc cccgattaca agaagctgtc
     2941 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gtctggtgac
     3001 cgtgacccag gactcctccc tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg
     3061 caccaacttc ccccccgacg gccccgtaat gcagaaaaag accatgggct gggaggcctc
     3121 caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccacc aggccctgaa
     3181 gctgaaggac ggcggccact acctggtgga gttcaagacc atctacatgg ccaagaagcc
     3241 cgtgcaactg cccggctact actacgtgga caccaagctg gacatcacct cccacaacga
     3301 ggactacacc atcgtggaac agtacgagcg ctccgagggc cgccaccacc tgttcctgta
     3361 cggcatggac gagctgtaca agtaagtcga ctaaaactga aacatccttc aagtctgtag
     3421 ttttgctgca gagaagaaaa tagttttcat ttttgagttt ttaacagaag tataaaacct
     3481 gacattgtca tagctcctgt ccttgtgttc tctctcctac acttaaaaag aaatacatac
     3541 tgactagtta tgtgagatgc taaaagttac aacattgcag ttactgatac aaatggtgtt
     3601 aactagaaat attaaagcat tctaaagtct ttcttatttc tcatatattc ttcagggggt
     3661 ttagatatgt ttaatgttta aatgccaagt cttactatag tgtttattga tcttataaaa
     3721 caagtatcta ggatatgata cacttttggt caaaaattac tgggtctcat ttttacttga
     3781 gcctttaaga cagtagtgtg tcactataat gtgatagcat gatctttaag aaacagtaga
     3841 agacttaagc ctttcaaagt gattttgatt tttagatcat cagatgtatg atgaaaatgg
     3901 ttaaatgttt gtgacatgag cgttgtactc aatggcataa caatgtttgt ttttataata
     3961 tacactcttt ccttgaaata aaggaagaaa cacttttccc ttaagttttc catcagcact
     4021 ttaaattttt ctttgttcag tatcctagcc ccaactcctg tatttttgaa actttgtttt
     4081 tttgtgtgtc ttttgtaacc aatgtagtga tcaaagtttc acataaatta tttatgtata
     4141 gtatacatta ctacacagat gtggcaatga aaatcaatct taagactcaa acccatttat
     4201 ccacaaaaaa cagtgctgct tactcttgcc ataagttttg atttatatct taagatcaat
     4261 ggaaagatcc ctgtgtttct tcattttcca ttatatatct taaacttact atgctactat
     4321 gcctagaata tacgaagttc ttttcttttt tttttttttt ttttttgaga cagagtcttg
     4381 ctcttatcac ccaggctgga gtacaatggt gcaatcttgg ctcactgcag cctctgcctc
     4441 ctgtgttcaa gcagttcttc tgtcttagcc tcctaattag ctgggactac agatgtgcac
     4501 catcatgcct ggctaagtat tatattttta gtagagatgg gctttcacca cgttggccag
     4561 gctggtctca aactcttgac ctcatgatcc gcccgccctg gcctctcaaa gtgctaagat
     4621 tacaggcata agccactgcg cctggcctga agttcttaat aaattgttaa atgaacgaac
     4681 ccaaatgttt aacacctaat gagttgtaaa taatggggag acaaaaatga ttggtattgt
     4741 tactgttcac ctaagcaatc caaaaatgac atgtccctga tgtatatctg gttaaggaat
     4801 ttagttaata aatgaggcat cacatatgaa aaaagagggc atatcaacag atagtgttag
     4861 gataactgga tagatggcta catcttgcca tattcaaaat taacttcatg tgaattatga
     4921 agtcctttcc ccattgcttt tgtccagttt atcaaagatc agctggttgt agatgtgtgg
     4981 ctttgcttct aaggtcaagg gcgaattctg cagatatcca tcacactggc ggccgctcga
     5041 gcatgcatct agagggccca attcgcccta tagtgagtcg tattacaatt cactggccgt
     5101 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     5161 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     5221 acagttgcgc agcctatacg tacggcagtt taaggtttac acctataaaa gagagagccg
     5281 ttatcgtctg tttgtggatg tacagagtga tattattgac acgccggggc gacggatggt
     5341 gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac tttacccggt
     5401 ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca gtgtgccggt
     5461 ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc
     5521 cattaacctg atgttctggg gaatataaat gtcaggcatg agattatcaa aaaggatctt
     5581 cacctagatc cttttcacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa
     5641 tgtcagctac tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc
     5701 ttgcagtggg cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc
     5761 ggaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat
     5821 ggctttctcg ccgccaagga tctgatggcg caggggatca agctctgatc aagagacagg
     5881 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5941 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     6001 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     6061 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     6121 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     6181 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     6241 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     6301 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     6361 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     6421 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     6481 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     6541 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     6601 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     6661 cttctatcgc cttcttgacg agttcttctg aattattaac gcttacaatt tcctgatgcg
     6721 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacaggtgg cacttttcgg
     6781 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6841 ctcatgagac aataaccctg ataaatgctt caataatagc acgtgaggag ggccaccatg
     6901 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag
     6961 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg
     7021 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac
     7081 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc
     7141 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg
     7201 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag
     7261 gagcaggact gacacgtgct aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7321 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7381 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7441 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7501 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     7561 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7621 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7681 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7741 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7801 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7861 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7921 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7981 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct gggcttttgc
     8041 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     8101 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     8161 gtgagcgagg aagcggaag
//
