LOCUS       sequence-266377-        4137 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 4137)
  AUTHORS   Gokirmak T, Campanale JP, Shipp LE, Moy GW, Tao H, Hamdoun A
  TITLE     Localization and Substrate Selectivity of Sea Urchin Multidrug (MDR)
            Efflux Transporters.
  JOURNAL   J Biol Chem. 2012 Nov 2.
  PUBMED    23124201
REFERENCE   2  (bases 1 to 4137)
  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: "J Biol 
            Chem. 2012 Nov 2."
COMMENT     Sequence Label: pCS2+8
FEATURES             Location/Qualifiers
     source          1..4137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        55..1039
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1074..1092
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1074..1091
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    1228..1362
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1252..1271)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1306..1325
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1475..1495)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1475..1493)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1514..1530)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1514..1530)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1527..1549)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1538..1554
                     /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(1562..1568,1569..1586,1587..1592))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1607..1628
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1745..1762)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1916..2504)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1996..2015)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2675..3466,3467..3535))
                     /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     3298..3317
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3536..3640)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3666..4121)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3803..3824)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4015..4034
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagtcgac
       61 cgccaattca atatggcgta tatggactca tgccaattca atatggtgga tctggacctg
      121 tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc
      181 agccaattca atatggcgga cttggcacca tgccaattca atatggcgga cctggcactg
      241 tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct
      301 gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat attggctata
      361 tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc
      421 aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat
      481 tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata
      541 ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga
      601 cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt
      661 aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca
      721 atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg
      781 gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta
      841 ctattggaag tacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat
      901 ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca
      961 aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag
     1021 caatgctcgt ttagggaacc gccattctgc ctggggacgt cggagcaagc ttgatttagg
     1081 tgacactata gaatacaagc tacttgttct ttttgcagga tccactagtg gcgcgccatt
     1141 aattaaaggc cggccagcga tcgccggaca tcgattggaa ttcaaggcct ctcgagcctc
     1201 tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt
     1261 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     1321 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     1381 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc ggccgcggcg
     1441 ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct
     1501 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     1561 acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     1621 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     1681 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     1741 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     1801 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     1861 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1921 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1981 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2041 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2101 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2161 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2221 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2281 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2341 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2401 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2461 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2521 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2581 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2641 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2701 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     2761 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     2821 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     2881 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     2941 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3001 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3061 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3121 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3181 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3241 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3301 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3361 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3421 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3481 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3541 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3601 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3661 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3721 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3781 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3841 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3901 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3961 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     4021 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     4081 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcag
//
