LOCUS       sequence-142911-        4854 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 4854)
  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 4854)
  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+8CeGFP
FEATURES             Location/Qualifiers
     source          1..4854
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(162..750)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(242..261)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(921..1712,1713..1781))
                     /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     1544..1563
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1782..1886)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1912..2367)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2049..2070)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2261..2280
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2438..3422
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        3457..3475
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3457..3474
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             3561..4274
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3591..3619)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4214..4234
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    4328..4462
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4352..4371)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4406..4425
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4575..4595)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4575..4593)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4614..4630)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4614..4630)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4627..4649)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4638..4654
                     /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(4662..4668,4669..4686,4687..4692))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4707..4728
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(join(4845..4854,1..8))
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
       61 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
      121 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      181 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      241 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      301 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      361 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      421 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      481 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      541 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      601 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      661 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      721 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      781 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      841 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      901 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
      961 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1021 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     1081 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1141 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1201 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1261 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1321 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1381 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1441 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1501 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1561 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1621 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1681 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1741 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1801 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1861 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     1921 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     1981 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     2041 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2101 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     2161 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2221 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2281 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2341 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     2401 aagggcgatc ggtgcgggcc tcttcgctat tacgccagtc gaccgccaat tcaatatggc
     2461 gtatatggac tcatgccaat tcaatatggt ggatctggac ctgtgccaat tcaatatggc
     2521 gtatatggac tcgtgccaat tcaatatggt ggatctggac cccagccaat tcaatatggc
     2581 ggacttggca ccatgccaat tcaatatggc ggacctggca ctgtgccaac tggggagggg
     2641 tctacttggc acggtgccaa gtttgaggag gggtcttggc cctgtgccaa gtccgccata
     2701 ttgaattggc atggtgccaa taatggcggc catattggct atatgccagg atcaatatat
     2761 aggcaatatc caatatggcc ctatgccaat atggctattg gccaggttca atactatgta
     2821 ttggccctat gccatatagt attccatata tgggttttcc tattgacgta gatagcccct
     2881 cccaatgggc ggtcccatat accatatatg gggcttccta ataccgccca tagccactcc
     2941 cccattgacg tcaatggtct ctatatatgg tctttcctat tgacgtcata tgggcggtcc
     3001 tattgacgta tatggcgcct cccccattga cgtcaattac ggtaaatggc ccgcctggct
     3061 caatgcccat tgacgtcaat aggaccaccc accattgacg tcaatgggat ggctcattgc
     3121 ccattcatat ccgttctcac gccccctatt gacgtcaatg acggtaaatg gcccacttgg
     3181 cagtacatca atatctatta atagtaactt ggcaagtaca ttactattgg aagtacgcca
     3241 gggtacattg gcagtactcc cattgacgtc aatggcggta aatggcccgc gatggctgcc
     3301 aagtacatcc ccattgacgt caatggggag gggcaatgac gcaaatgggc gttccattga
     3361 cgtaaatggg cggtaggcgt gcctaatggg aggtctatat aagcaatgct cgtttaggga
     3421 accgccattc tgcctgggga cgtcggagca agcttgattt aggtgacact atagaataca
     3481 agctacttgt tctttttgca ggatccacta gtggcgcgcc attaattaaa ggccggccag
     3541 cgatcgccgg acatcgattg atgagtaaag gagaagaact tttcactgga gttgtcccaa
     3601 ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg
     3661 aaggtgatgc aacatacgga aaacttaccc ttaaatttat ttgcactact ggaaaactac
     3721 ctgttccatg gccaacactt gtcactactc tcacttatgg tgttcaatgc ttttcaagat
     3781 acccagatca tatgaaacgg catgactttt tcaagagtgc catgcccgaa ggttatgtac
     3841 aggaaagaac tatatttttc aaagatgacg ggaactacaa gacacgtgct gaagtcaagt
     3901 ttgaaggtga tacccttgtt aatagaatcg agttaaaagg tattgatttt aaagaagatg
     3961 gaaacattct tggacacaaa ttggaataca actataactc acacaatgta tacatcatgg
     4021 cagacaaaca aaagaatgga atcaaagtta acttcaaaat tagacacaac attgaagatg
     4081 gaagcgttca actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc
     4141 ttttaccaga caaccattac ctgtccacac aatctgccct ttcgaaagat cccaacgaaa
     4201 agagagacca catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg
     4261 atgaactata caaataagaa ttcaaggcct ctcgagcctc tagaactata gtgagtcgta
     4321 ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     4381 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     4441 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     4501 gggggaggtg tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt gggcccggta
     4561 cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     4621 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccgggagc
     4681 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4741 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4801 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cact
//
