LOCUS       sequence-45567-e        4618 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 4618)
  AUTHORS   Wang JW, Beck ES, McCabe BD
  TITLE     A modular toolset for recombination transgenesis and neurogenetic 
            analysis of Drosophila.
  JOURNAL   PLoS One. 2012;7(7):e42102. Epub 2012 Jul 25.
  PUBMED    22848718
REFERENCE   2  (bases 1 to 4618)
  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: "PLoS One.";
            date: "2012"; volume: "7(7)"; pages: "e42102. Epub 2012 Jul 25"
COMMENT     Sequence Label: pMartini Gate A R2-R1
FEATURES             Location/Qualifiers
     source          1..4618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(33..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(143..164)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     355..374
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    701..825
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(866..1171)
                     /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     complement(968..987)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1513..2172)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2087..2106
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(2226..2232,2233..2250,2251..2256))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(2281..2405)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2472..2488)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2472..2488)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2485..2507)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2496..2512
                     /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(2520..2526,2527..2544,2545..2550))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2565..2586
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2703..2720)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2874..3462)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2954..2973)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3633..4424,4425..4493))
                     /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     4256..4275
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4494..4598)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
       61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg agcggccgcc ggcctcgagg gccggatctg caggtcgacc
      661 agctggccaa gcttcttcta gaggtaccgc atgcgatatc accactttgt acaagaaagc
      721 tgaacgagaa acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa
      781 acagactaca taatactgta aaacacaaca tatccagtca ctatggtcga cctgcagact
      841 ggctgtgtat aagggagcct gacatttata ttccccagaa catcaggtta atggcgtttt
      901 tgatgtcatt ttcgcggtgg ctgagatcag ccacttcttc cccgataacg gagaccggca
      961 cactggccat atcggtggtc atcatgcgcc agctttcatc cccgatatgc accaccgggt
     1021 aaagttcacg ggagacttta tctgacagca gacgtgcact ggccaggggg atcaccatcc
     1081 gtcgcccggg cgtgtcaata atatcactct gtacatccac aaacagacga taacggctct
     1141 ctcttttata ggtgtaaacc ttaaactgca tttcaccagc ccctgttctc gtcagcaaaa
     1201 gagccgttca tttcaataaa ccgggcgacc tcagccatcc cttcctgatt ttccgctttc
     1261 cagcgttcgg cacgcagacg acgggcttca ttctgcatgg ttgtgcttac cagaccggag
     1321 atattgacat catatatgcc ttgagcaact gatagctgtc gctgtcaact gtcactgtaa
     1381 tacgctgctt catagcatac ctctttttga catacttcgg gtatacatat cagtatatat
     1441 tcttataccg caaaaatcag cgcgcaaata cgcatactgt tatctggctt ttagtaagcc
     1501 ggatccacgc gtttacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag
     1561 cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat
     1621 cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt
     1681 gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac
     1741 gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc
     1801 cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag
     1861 cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca
     1921 tatcaccagc tcaccgtctt tcattgccat acggaattcc ggatgagcat tcatcaggcg
     1981 ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa
     2041 aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa
     2101 tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat
     2161 ttttttctcc attttagctt ccttagctcc tgaaaatctc gacggatcct aactcaaaat
     2221 ccacacatta tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgcggccgc
     2281 catagtgact ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc
     2341 taatttaata tattgatatt tatatcattt tacgtttctc gttcagcttt tttgtacaaa
     2401 cttgtgatat cgagctctcc cgggaattcg gatccggccc tcgaggccgg cggccgcttg
     2461 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     2521 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     2581 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     2641 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     2701 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2761 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2821 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2881 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2941 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3001 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3061 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3121 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3181 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3241 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3301 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3361 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3421 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3481 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3541 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3601 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3661 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3721 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3781 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3841 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3901 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3961 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4021 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4081 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4141 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4201 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4261 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4321 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4381 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4441 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4501 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4561 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgc
//
