LOCUS Exported 4198 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBS-ermGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4198) AUTHORS Lizier M, Sarra PG, Cauda R, Lucchini F TITLE Comparison of expression vectors in Lactobacillus reuteri strains. JOURNAL FEMS Microbiol Lett. 2010 Jul 1. 308(1):8-15. PUBMED 20455948 REFERENCE 2 (bases 1 to 4198) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4198 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(8..24) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(9..25) /label=pBluescriptKS /note="For pBluescript vector" primer_bind complement(54..74) /label=T3 /note="T3 promoter, forward primer" promoter complement(54..72) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(93..109) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(93..109) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(106..128) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 117..133 /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(141..171) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 186..207 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(324..341) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(495..1083) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(575..594) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1254..2114) /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 1877..1896 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2115..2219) /gene="bla" /label=AmpR promoter rep_origin complement(2245..2700) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2382..2403) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2594..2613 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2827..2849 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2841..2858 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2842..2858 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 2868..2887 /label=T7 /note="T7 promoter, forward primer" promoter 2868..2886 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 2919..2935 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 2919..2935 /label=pBluescriptSK /note="For pBluescript vector" CDS complement(2937..3653) /codon_start=1 /gene="mgfp5" /product="GFP with folding enhancement mutations" /label=mgfp5 /note="suitable for expression in plants due to removal of a cryptic intron" /translation="MSKGEELFTGVVPILVELDGDVNGYKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA NFKTRHNIEDGGVQLADHYQQNTPIGDDPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" primer_bind complement(2980..3000) /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" primer_bind 3595..3623 /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" ORIGIN 1 agcttatcga taccgtcgac ctcgaggggg ggcccggtac ccagcttttg ttccctttag 61 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt 121 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt 181 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg 241 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg 301 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 361 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 421 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 481 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 541 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 601 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 661 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg 721 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 781 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 841 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 901 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg 961 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 1021 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 1081 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 1141 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 1201 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 1261 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 1321 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 1381 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 1441 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 1501 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 1561 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 1621 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 1681 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 1741 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 1801 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 1861 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 1921 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 1981 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 2041 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 2101 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 2161 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 2221 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt 2281 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt 2341 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc 2401 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg 2461 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac 2521 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg 2581 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag 2641 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt 2701 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc 2761 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag 2821 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac 2881 tatagggcga attggagctc caccgcggtg gcggccgctc tagaactagt ggatccttat 2941 ttgtatagtt catccatgcc atgtgtaatc ccagcagccg ttacaaactc aagaaggacc 3001 atgtggtctc tcttttcgtt gggatctttc gaaagggcag attgtgtgga caggtaatgg 3061 ttgtctggta aaaggacagg gtcatcgcca attggagtat tttgttgata atgatcagcg 3121 agttgcacgc cgccgtcttc gatgttgtgg cgggtcttga agttggcttt gatgccgttc 3181 ttttgcttgt cggccatgat gtatacgttg tgggagttgt agttgtattc caacttgtgg 3241 ccgaggatgt ttccgtcctc cttgaaatcg attcccttaa gctcgatcct gttgacgagg 3301 gtgtctccct caaacttgac ttcagcacgt gtcttgtagt tcccgtcgtc cttgaagaag 3361 atggtcctct cctgcacgta tccctcaggc atggcgctct tgaagaagtc gtgccgcttc 3421 atatgatctg gatatcttga aaagcattga acaccataag tgagagtagt gacaagtgtt 3481 ggccatggaa caggtagttt tccagtagtg caaataaatt taagggtaag ttttccgtat 3541 gttgcatcac cttcaccctc tccactgaca gaaaatttgt acccattaac atcaccatct 3601 aattcaacaa gaattgggac aactccagtg aaaagttctt ctcctttact catgaattca 3661 ctccttctta attacaaatt tttagcatct aatttaactt caattcctat tatacaaaat 3721 tttaagatac tgcactatca acacactctt aagtttgctt ctaagtctta tttccataac 3781 ttcttttacg tttccgccat tctttgctgt ttcgattttt atgatatggt gcaagtcagc 3841 acgaacacga accgtcttat ctcccattat atcttttttt gcactgattg gtgtatcatt 3901 tcgtttttct ttttgtgcgc ttcttgataa aagggatagt aattcattcc tggttgcaaa 3961 ttttgaaaac cgctacggat cacatctttt tctaaactat tgatccatag tcttttatac 4021 gttttatctt tagaaaaggc atttgcttta tgaatgatcg accaggcaat gttttcgcct 4081 tctctgtcgc tatctgttgc gacaatgatt gtatttgctt gttttaaaag ttcagcaaca 4141 attttaaact gctttttttt atctgttgcc acttcaaaat cgtatcgatt ctagacta //