LOCUS Exported 4854 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCS2+8CeGFP 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4854 /organism="synthetic DNA construct" /mol_type="other DNA" 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(921..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 2445..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..4277 /codon_start=1 /product="Aequoria victoria green fluorescent protein" /label=GFP /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" promoter complement(4306..4323) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" 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(4662..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(4845..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 //