LOCUS       sequence-7445-ex        6530 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6530)
  AUTHORS   Villefranc JA, Amigo J, Lawson ND
  TITLE     Gateway compatible vectors for analysis of gene function in the 
            zebrafish.
  JOURNAL   Dev Dyn. 2007 Nov . 236(11):3077-87.
  PUBMED    17948311
REFERENCE   2  (bases 1 to 6530)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Dev Dyn. 
            2007 Nov . 236(11):3077-87."
COMMENT     Sequence Label: 362 pCS Cherry DEST
FEATURES             Location/Qualifiers
     source          1..6530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      109..118
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             115..822
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(260..278)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     528..547
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     722..745
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    835..959
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(984..989,990..1007,1008..1014)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1068..1727
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1134..1153)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2069..2374
                     /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="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2253..2272
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2415..2539)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    2579..2713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2603..2622)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2657..2676
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2826..2846)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2826..2844)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2865..2881)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2865..2881)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2878..2900)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2889..2905
                     /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)."
     promoter        complement(join(2913..2919,2920..2937,2938..2943))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2958..2979
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3096..3113)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3267..3855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3347..3366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4026..4817,4818..4886))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4649..4668
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4887..4991)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5017..5472)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5154..5175)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5366..5385
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5546..6530
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattca aggctagcgc caccatggtg
      121 agcaagggcg aggaggacaa catggccatc atcaaggagt tcatgcgctt caaggtgcac
      181 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
      241 tacgagggca cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc
      301 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
      361 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
      421 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
      481 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
      541 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
      601 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgccgaggtc
      661 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
      721 aagctggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgcgcc
      781 gagggccgcc actccaccgg cggcatggac gagctgtaca agagatctcg aatcacaagt
      841 ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa atatcaatat attaaattag
      901 attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgg
      961 cggccgcatt aggcacccca ggctttacac tttatgcttc cggctcgtat aatgtgtgga
     1021 ttttgagtta ggatccgtcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
     1081 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
     1141 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
     1201 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     1261 gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat
     1321 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
     1381 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
     1441 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
     1501 tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
     1561 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
     1621 tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc
     1681 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaaacg cgtggatccg
     1741 gcttactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa
     1801 tatatactga tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat
     1861 tacagtgaca gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc
     1921 tccggtctgg taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg
     1981 aaagcggaaa atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct
     2041 tttgctgacg agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga
     2101 gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg
     2161 gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta
     2221 cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt
     2281 gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa
     2341 aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca
     2401 gtctgcaggt cgaccatagt gactggatat gttgtgtttt acagcattat gtagtctgtt
     2461 ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag
     2521 ctttcttgta caaagtggtg attcgagcct ctagaactat agtgagtcgt attacgtaga
     2581 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2641 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2701 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     2761 gtgggaggtt ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt
     2821 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     2881 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     2941 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3001 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3061 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3121 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3181 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3241 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3301 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3361 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3421 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3481 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3541 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3601 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3661 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     3721 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3781 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3841 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3901 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3961 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4021 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4081 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4141 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4201 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4261 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4321 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4381 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4441 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4501 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4561 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4621 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4681 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4741 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4801 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4861 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4921 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4981 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     5041 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     5101 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     5161 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     5221 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     5281 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     5341 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     5401 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     5461 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     5521 gggcctcttc gctattacgc cagtcgacca tagccaattc aatatggcgt atatggactc
     5581 atgccaattc aatatggtgg atctggacct gtgccaattc aatatggcgt atatggactc
     5641 gtgccaattc aatatggtgg atctggaccc cagccaattc aatatggcgg acttggcacc
     5701 atgccaattc aatatggcgg acttggcact gtgccaactg gggaggggtc tacttggcac
     5761 ggtgccaagt ttgaggaggg gtcttggccc tgtgccaagt ccgccatatt gaattggcat
     5821 ggtgccaata atggcggcca tattggctat atgccaggat caatatatag gcaatatcca
     5881 atatggccct atgccaatat ggctattggc caggttcaat actatgtatt ggccctatgc
     5941 catatagtat tccatatatg ggttttccta ttgacgtaga tagcccctcc caatgggcgg
     6001 tcccatatac catatatggg gcttcctaat accgcccata gccactcccc cattgacgtc
     6061 aatggtctct atatatggtc tttcctattg acgtcatatg ggcggtccta ttgacgtata
     6121 tggcgcctcc cccattgacg tcaattacgg taaatggccc gcctggctca atgcccattg
     6181 acgtcaatag gaccacccac cattgacgtc aatgggatgg ctcattgccc attcatatcc
     6241 gttctcacgc cccctattga cgtcaatgac ggtaaatggc ccacttggca gtacatcaat
     6301 atctattaat agtaacttgg caagtacatt actattggaa ggacgccagg gtacattggc
     6361 agtactccca ttgacgtcaa tggcggtaaa tggcccgcga tggctgccaa gtacatcccc
     6421 attgacgtca atggggaggg gcaatgacgc aaatgggcgt tccattgacg taaatgggcg
     6481 gtaggcgtgc ctaatgggag gtctatataa gcaatgctcg tttagggaac
//
