LOCUS       sequence-10309-e        5808 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 5808)
  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 5808)
  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: pCSDest
FEATURES             Location/Qualifiers
     source          1..5808
                     /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"
     protein_bind    108..232
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(257..262,263..280,281..287)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             341..1000
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(407..426)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1342..1647
                     /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     1526..1545
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1688..1812)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    1857..1991
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1881..1900)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1935..1954
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2104..2124)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2104..2122)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2143..2159)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2143..2159)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2156..2178)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2167..2183
                     /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(2191..2197,2198..2215,2216..2221))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2236..2257
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2374..2391)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2545..3133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2625..2644)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3304..4095,4096..4164))
                     /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     3927..3946
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4165..4269)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4295..4750)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4432..4453)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4644..4663
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4824..5808
                     /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 aggatcaaca agtttgtaca
      121 aaaaagctga acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc
      181 ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcatat tggcggccgc
      241 attaggcacc ccaggcttta cactttatgc ttccggctcg tataatgtgt ggattttgag
      301 ttaggatccg tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg
      361 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
      421 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
      481 cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
      541 gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag
      601 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag
      661 tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta
      721 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc
      781 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
      841 cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
      901 ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga
      961 attacaacag tactgcgatg agtggcaggg cggggcgtaa agatctggat ccggcttact
     1021 aaaagccaga taacagtatg cgtatttgcg cgctgatttt tgcggtataa gaatatatac
     1081 tgatatgtat acccgaagta tgtcaaaaag aggtatgcta tgaagcagcg tattacagtg
     1141 acagttgaca gcgacagcta tcagttgctc aaggcatata tgatgtcaat atctccggtc
     1201 tggtaagcac aaccatgcag aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg
     1261 aaaatcagga agggatggct gaggtcgccc ggtttattga aatgaacggc tcttttgctg
     1321 acgagaacag gggctggtga aatgcagttt aaggtttaca cctataaaag agagagccgt
     1381 tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg acggatggtg
     1441 atccccctgg ccagtgcacg tctgctgtca gataaagtct cccgtgaact ttacccggtg
     1501 gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggtc
     1561 tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc
     1621 attaacctga tgttctgggg aatataaatg tcaggctccc ttatacacag ccagtctgca
     1681 ggtcgaccat agtgactgga tatgttgtgt tttacagtat tatgtagtct gttttttatg
     1741 caaaatctaa tttaatatat tgatatttat atcattttac gtttctcgtt cagctttctt
     1801 gtacaaagtg gttgatcctc tcgagcctct agaactatag tgagtcgtat tacgtagatc
     1861 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     1921 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     1981 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     2041 gggaggtttt ttaattcgcg gccgcggcgc caatgcattg ggcccggtac ccagcttttg
     2101 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     2161 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     2221 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     2281 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     2341 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     2401 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2461 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2521 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2581 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2641 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2701 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2761 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2821 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2881 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2941 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     3001 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3061 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3121 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3181 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3241 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3301 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3361 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3421 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3481 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3541 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3601 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3661 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3721 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3781 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3841 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3901 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3961 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4021 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4081 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4141 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4201 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4261 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     4321 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4381 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4441 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4501 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4561 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4621 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4681 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4741 cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     4801 gcctcttcgc tattacgcca gtcgaccata gccaattcaa tatggcgtat atggactcat
     4861 gccaattcaa tatggtggat ctggacctgt gccaattcaa tatggcgtat atggactcgt
     4921 gccaattcaa tatggtggat ctggacccca gccaattcaa tatggcggac ttggcaccat
     4981 gccaattcaa tatggcggac ttggcactgt gccaactggg gaggggtcta cttggcacgg
     5041 tgccaagttt gaggaggggt cttggccctg tgccaagtcc gccatattga attggcatgg
     5101 tgccaataat ggcggccata ttggctatat gccaggatca atatataggc aatatccaat
     5161 atggccctat gccaatatgg ctattggcca ggttcaatac tatgtattgg ccctatgcca
     5221 tatagtattc catatatggg ttttcctatt gacgtagata gcccctccca atgggcggtc
     5281 ccatatacca tatatggggc ttcctaatac cgcccatagc cactccccca ttgacgtcaa
     5341 tggtctctat atatggtctt tcctattgac gtcatatggg cggtcctatt gacgtatatg
     5401 gcgcctcccc cattgacgtc aattacggta aatggcccgc ctggctcaat gcccattgac
     5461 gtcaatagga ccacccacca ttgacgtcaa tgggatggct cattgcccat tcatatccgt
     5521 tctcacgccc cctattgacg tcaatgacgg taaatggccc acttggcagt acatcaatat
     5581 ctattaatag taacttggca agtacattac tattggaagg acgccagggt acattggcag
     5641 tactcccatt gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt acatccccat
     5701 tgacgtcaat ggggaggggc aatgacgcaa atgggcgttc cattgacgta aatgggcggt
     5761 aggcgtgcct aatgggaggt ctatataagc aatgctcgtt tagggaac
//
