LOCUS       sequence-10318-e        6761 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 6761)
  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 6761)
  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: pCSCherryN5icd
FEATURES             Location/Qualifiers
     source          1..6761
                     /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..859
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2746..2770)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2810..2944
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2834..2853)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2888..2907
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3057..3077)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3057..3075)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3096..3112)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3096..3112)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3109..3131)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3120..3136
                     /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(3144..3150,3151..3168,3169..3174))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3189..3210
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3327..3344)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3498..4086)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3578..3597)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4257..5048,5049..5117))
                     /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     4880..4899
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5118..5222)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5248..5703)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5385..5406)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5597..5616
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5777..6761
                     /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 aagcaggctt gatggttggc atgttgattg cccgccgcaa gcgtgaacac
      901 agcacactct ggttccctga gggcttcttc ctcaaaaagg aaacaagcag taacaagaac
      961 cgcagagaac ctgtgggcca agattcactg ggcatgaaac acatgccaaa gacagtggaa
     1021 gagtctcttc tggctgatca cagtgaccag tggatagata cagactgccc agaggctaag
     1081 cgacttaagg tggaagaacc aagcattctg tcagatggtg aggatgcagt tgacagcaga
     1141 cagtggacac agcatcactt ggcagcagca gatatccgca tgccaccttc catggcactt
     1201 acaccaccac aaggagagtt tgacagtgac tgcatggatg ttaatgtcag aggccctgat
     1261 ggcttcacac ctctgatgct ggcatctttc tgtggaggag ggcttgagcc agaggtgacc
     1321 gaagatgatg actcagatga atcctcagcc aacatcatat ctgacctcat ttaccaggga
     1381 gcatcacttg cagctcaaac tgaccgcact ggagagactg ctctgcacct ggctgcacgc
     1441 tatgcccggg cagatgcagc taaaagactc ctggatgctg gggctgatgc aaatgcacag
     1501 gataacacag gtcgctcacc tttgcatgca gctgtggcag ctgatgcaca gggagttttc
     1561 cagattctga tcaggaaccg tgccacagac ttggacgcgc gcatgtatga tggctccact
     1621 gctctgatac ttgcagcccg tctggcagta gagggcatgg tggaagagtt gatcacctgc
     1681 catgctgatg ttaatgctgt tgatgaaatc ggaaagtcag cattgcactg ggcagcagca
     1741 gtcaataacg ttgaagctac tattgccttg ttgaagaatg gtgccaataa agatatgcaa
     1801 gacctcaagg aggagactcc gctgttcttg gctgccaggg aaggcagctg tgaggcagta
     1861 aaggtgttgt tggctcactt tgccaatagg gaaataacag atcacatgga cagactccct
     1921 cgagatattg cccaggagcg catgcatcat gacattgtgc agctactgga tgaatataac
     1981 actgtgcgaa gtccacaggg tcatgcaggg gcaggtcacc acctctctgg tagccacaca
     2041 ctctcaccgc tcatgtgccc tcccagcaac ttcctccaag ggctgaaaag tacccctcag
     2101 ggtaagaaga accgtcgccc gggagctaaa ggcattggtg gacagcacgc ttccggcctg
     2161 aaagattcag ccaaaggacg caacaaacgg ctgactctag acatgcagag tgctttgctg
     2221 gagagctctg tcacgctctc ccctgttgac tcactggatt caccacgagg aggcgccagt
     2281 aacgctggct atgtcaccaa tccaacctca ccagctgcca tgccctctcc cggccttttc
     2341 cactcctcta tgtctgtccc taataccccc atggtgcaca gcagtatcct ggacagcacc
     2401 agcccttttt ctgtctccct ggcacagttg agtgacctgg gagatggtgg gctctcaatg
     2461 caggggcgtg tgggcatgca aggagggggt gtcaacccag gcccccataa ctacgtgatg
     2521 aacgccggtc aaatgagcct tagtatgggc atggtaagtc cagtaagcgt gccgtttgac
     2581 tggcataacc gcatgcctcc atcttctcag tgcagtcagc caatgatcag catggtgcat
     2641 ccagtaagta gtcaagcagg catgctccca caaacaccaa ccctgcaaag cagcatgcta
     2701 ttgcaccagc aggtgttccg taatgcccag cagcccatcc tgcagaccca gctttcttgt
     2761 acaaagtggt gattcgagcc tctagaacta tagtgagtcg tattacgtag atccagacat
     2821 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2881 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2941 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     3001 tttttaattc gcggccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt
     3061 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3121 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     3181 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3241 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3301 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3361 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3421 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3481 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3541 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3601 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3661 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3721 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3781 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3841 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3901 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     3961 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4021 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4081 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4141 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4201 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4261 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4321 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4381 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4441 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4501 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4561 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4621 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4681 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4741 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4801 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4861 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4921 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4981 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5041 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5101 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5161 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5221 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     5281 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     5341 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     5401 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     5461 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     5521 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     5581 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     5641 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     5701 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     5761 cgctattacg ccagtcgacc atagccaatt caatatggcg tatatggact catgccaatt
     5821 caatatggtg gatctggacc tgtgccaatt caatatggcg tatatggact cgtgccaatt
     5881 caatatggtg gatctggacc ccagccaatt caatatggcg gacttggcac catgccaatt
     5941 caatatggcg gacttggcac tgtgccaact ggggaggggt ctacttggca cggtgccaag
     6001 tttgaggagg ggtcttggcc ctgtgccaag tccgccatat tgaattggca tggtgccaat
     6061 aatggcggcc atattggcta tatgccagga tcaatatata ggcaatatcc aatatggccc
     6121 tatgccaata tggctattgg ccaggttcaa tactatgtat tggccctatg ccatatagta
     6181 ttccatatat gggttttcct attgacgtag atagcccctc ccaatgggcg gtcccatata
     6241 ccatatatgg ggcttcctaa taccgcccat agccactccc ccattgacgt caatggtctc
     6301 tatatatggt ctttcctatt gacgtcatat gggcggtcct attgacgtat atggcgcctc
     6361 ccccattgac gtcaattacg gtaaatggcc cgcctggctc aatgcccatt gacgtcaata
     6421 ggaccaccca ccattgacgt caatgggatg gctcattgcc cattcatatc cgttctcacg
     6481 ccccctattg acgtcaatga cggtaaatgg cccacttggc agtacatcaa tatctattaa
     6541 tagtaacttg gcaagtacat tactattgga aggacgccag ggtacattgg cagtactccc
     6601 attgacgtca atggcggtaa atggcccgcg atggctgcca agtacatccc cattgacgtc
     6661 aatggggagg ggcaatgacg caaatgggcg ttccattgac gtaaatgggc ggtaggcgtg
     6721 cctaatggga ggtctatata agcaatgctc gtttagggaa c
//
