LOCUS       sequence-142449-        4848 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 4848)
  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 4848)
  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+8CmCherry
FEATURES             Location/Qualifiers
     source          1..4848
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     240..260
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    373..507
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(410..429)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     464..483
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(558..1268)
                     /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(836..855)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1105..1123
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     promoter        complement(1354..1372)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1355..1372)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(1407..2391)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     rep_origin      2462..2917
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2549..2568)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2759..2780
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2943..3047
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3048..3116,3117..3908)
                     /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     complement(3266..3285)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4079..4667
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4568..4587
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4821..4838
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgctccc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
      241 caattaaccc tcactaaagg gaacaaaagc tgggtaccgg gcccaatgca ttggcgccgc
      301 ggccgcgaat taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
      361 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
      421 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
      481 atcaatgtat cttatcatgt ctggatctac gtaatacgac tcactatagt tctagaggct
      541 cgagaggcct tgaattctta cttgtacagc tcgtccatgc cgccggtgga gtggcggccc
      601 tcggcgcgtt cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt gatgtccaac
      661 ttgatgttga cgttgtaggc gccgggcagc tgcacgggct tcttggcctt gtaggtggtc
      721 ttgacctcag cgtcgtagtg gccgccgtcc ttcagcttca gcctctgctt gatctcgccc
      781 ttcagggcgc cgtcctcggg gtacatccgc tcggaggagg cctcccagcc catggtcttc
      841 ttctgcatta cggggccgtc ggaggggaag ttggtgccgc gcagcttcac cttgtagatg
      901 aactcgccgt cctgcaggga ggagtcctgg gtcacggtca ccacgccgcc gtcctcgaag
      961 ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcaagta gtcggggatg
     1021 tcggcggggt gcttcacgta ggccttggag ccgtacatga actgagggga caggatgtcc
     1081 caggcgaagg gcagggggcc acccttggtc accttcagct tggcggtctg ggtgccctcg
     1141 taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcac ggagccctcc
     1201 atgtgcacct tgaagcgcat gaactccttg atgatggcca tgttatcctc ctcgcccttg
     1261 ctcaccatca atcgatgtcc ggcgatcgct ggccggcctt taattaatgg cgcgccacta
     1321 gtggatcctg caaaaagaac aagtagcttg tattctatag tgtcacctaa atcaagcttg
     1381 ctccgacgtc cccaggcaga atggcggttc cctaaacgag cattgcttat atagacctcc
     1441 cattaggcac gcctaccgcc catttacgtc aatggaacgc ccatttgcgt cattgcccct
     1501 ccccattgac gtcaatgggg atgtacttgg cagccatcgc gggccattta ccgccattga
     1561 cgtcaatggg agtactgcca atgtaccctg gcgtacttcc aatagtaatg tacttgccaa
     1621 gttactatta atagatattg atgtactgcc aagtgggcca tttaccgtca ttgacgtcaa
     1681 tagggggcgt gagaacggat atgaatgggc aatgagccat cccattgacg tcaatggtgg
     1741 gtggtcctat tgacgtcaat gggcattgag ccaggcgggc catttaccgt aattgacgtc
     1801 aatgggggag gcgccatata cgtcaatagg accgcccata tgacgtcaat aggaaagacc
     1861 atatatagag accattgacg tcaatggggg agtggctatg ggcggtatta ggaagcccca
     1921 tatatggtat atgggaccgc ccattgggag gggctatcta cgtcaatagg aaaacccata
     1981 tatggaatac tatatggcat agggccaata catagtattg aacctggcca atagccatat
     2041 tggcataggg ccatattgga tattgcctat atattgatcc tggcatatag ccaatatggc
     2101 cgccattatt ggcaccatgc caattcaata tggcggactt ggcacagggc caagacccct
     2161 cctcaaactt ggcaccgtgc caagtagacc cctccccagt tggcacagtg ccaggtccgc
     2221 catattgaat tggcatggtg ccaagtccgc catattgaat tggctggggt ccagatccac
     2281 catattgaat tggcacgagt ccatatacgc catattgaat tggcacaggt ccagatccac
     2341 catattgaat tggcatgagt ccatatacgc catattgaat tggcggtcga ctggcgtaat
     2401 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     2461 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2521 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2581 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2641 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2701 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2761 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2821 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2881 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     2941 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3001 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3061 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3121 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3181 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3241 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3301 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3361 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3421 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3481 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3541 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3601 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3661 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3721 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3781 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3841 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3901 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     3961 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4021 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4081 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4141 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4201 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4261 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4321 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4381 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4441 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4501 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4561 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4621 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4681 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4741 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4801 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
