LOCUS       Exported                4666 bp ds-DNA     circular SYN 22-AUG-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Addgene207869_p0201_pRset_OppA
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 4666)
  AUTHORS   TANTAMA LAB
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Aug 22, 2023 from SnapGene 3.2.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4666
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     rep_origin      10..465
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        491..595
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             596..1456
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1627..2215
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        2471..2489
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2535..2541
                     /note="RBS"
                     /note="ribosome binding site"
     CDS             2551..2553
                     /product="start codon"
                     /note="ATG"
     CDS             2563..2580
                     /product="6xHis affinity tag"
                     /note="6xHis"
     CDS             2584..2616
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /note="T7 tag (gene 10 leader)"
                     /note="promotes efficient translation in E. coli"
     CDS             2620..2643
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /note="Xpress(TM) tag"
     CDS             2653..4455
                     /codon_start=1
                     /note="L. lactis cremoris MG1363 OppA C23S"
                     /translation="MNKLKVTLLASSVVLAATLLSASGSNQSSSTSTKKLKAGNFDVAY
                     QNPDKAIKGGNLKVAYQSDSPMKAQWLSGLSNDATFATMSGPGGGQDGLFFTDSGFKFI
                     KGGAADVALDKESKTATITLRKDLKWSDGSEVTAKDYEFTYETIANPAYGSDRWTDSLA
                     NIVGLSDYHTGKAKTISGITFPDGENGKVIKVQFKEMKPGMTQSGNGYFLETVAPYQYL
                     KDVAPKDLASSPKTTTKPLVTGPFKPENVVAGESIKYVPNPYYWGEKPKLNSITYEVVS
                     TAKSVAALSSSKYDIINGMVSSQYKQVKNLKGYKVLGQQAMYISLMYYNLGHYDAKNSI
                     NVQDRKTPLQDQNVRQAIGYARNVAEVDNKFSNGLSTPANSLIPPIFKQFTSSSVKGYE
                     KQDLDKANKLLDEDGWKLNKSTGYREKDGKELSLVYAARVGDANAETIAQNYIQQWKKI
                     GVKVSLYNGKLMEFNSWVDHMTTPPGANDWDITDGSWSLASEPSQQDLFSAAAPYNFGH
                     FNDSEITKDLNDIDSAKSENPTYRKAAFVKYQEDMNKKAYVIPTNFMLNYTPVNKRVVG
                     MTLDYGAMNTWSEIGVSSAKLATK"
     terminator      4532..4579
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
       61 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
      121 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
      181 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
      241 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
      301 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
      361 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
      421 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
      481 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
      541 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
      601 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
      661 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
      721 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
      781 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
      841 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
      901 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
      961 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1021 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1081 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     1141 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     1201 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     1261 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     1321 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     1381 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     1441 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     1501 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     1561 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     1621 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     1681 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     1741 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     1801 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     1861 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     1921 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     1981 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2041 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2101 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2161 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2221 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     2281 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     2341 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     2401 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca ggatctcgat
     2461 cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc tagaaataat
     2521 tttgtttaac tttaagaagg agatatacat atgcggggtt ctcatcatca tcatcatcat
     2581 ggtatggcta gcatgactgg tggacagcaa atgggtcggg atctgtacga cgatgacgat
     2641 aaggatccca ccatgaataa gttgaaagtc accctccttg ctagttcagt cgtgcttgct
     2701 gctacactgc tgagcgctag tggttctaat caaagctcat ccacttccac caagaagctc
     2761 aaggcaggta actttgatgt ggcataccag aatcctgaca aagctatcaa gggaggcaat
     2821 ttgaaggtag cttaccaaag cgactcccct atgaaggcac agtggctgtc tggtctgtct
     2881 aatgacgcta ctttcgcaac tatgtctggc ccaggtgggg gacaggacgg actgtttttt
     2941 acagacagcg ggtttaagtt catcaaggga ggggcagcag acgtagcact ggataaggag
     3001 tctaagaccg ccacaattac tctccgaaag gatctgaaat ggtccgacgg cagtgaggtc
     3061 acggccaaag actatgagtt tacctacgag accatcgcaa acccagctta tggctccgac
     3121 cgttggaccg actctcttgc taacattgta ggtctcagtg actaccatac aggcaaagct
     3181 aaaaccatct ccggtatcac tttccctgat ggagagaacg gcaaggtcat caaggtgcag
     3241 tttaaggaaa tgaaaccagg aatgacacaa agcggaaatg gctattttct ggagacggtc
     3301 gcaccatatc agtatcttaa ggatgtcgct ccaaaggacc tcgcttctag ccccaaaaca
     3361 acaacaaaac ctctcgtgac cggacccttc aagccagaga acgtggttgc cggcgaatct
     3421 atcaaatacg tgcctaatcc ttactattgg ggtgaaaagc ctaagctgaa cagcatcact
     3481 tacgaagttg tatctacagc taagtcagtg gcagccttga gttcttctaa gtacgatatt
     3541 atcaatggga tggtctcttc tcagtataag caggtgaaga atctgaaagg ttataaagta
     3601 ctcggtcagc aagccatgta tattagcctg atgtactata atctcggcca ttatgacgca
     3661 aagaactcta tcaatgttca ggatcgcaag actcccctcc aggatcagaa tgttcggcag
     3721 gccatcggat acgctcgcaa cgtcgctgaa gttgacaata agttctccaa cggattgagc
     3781 acacctgcta attctcttat tccccccata ttcaagcagt ttactagcag ctctgtgaaa
     3841 ggctatgaga aacaagacct ggacaaggca aacaagctgt tggatgagga tggttggaag
     3901 ctgaataagt ctacaggtta tcgggaaaag gacggcaagg aactctccct ggtgtacgct
     3961 gctcgtgtag gggatgccaa cgctgaaact atcgcacaga attacatcca gcaatggaag
     4021 aaaatcggtg tcaaggtgtc tctttacaac ggcaaactga tggagttcaa tagttgggta
     4081 gaccacatga ccacaccacc aggggccaac gattgggaca tcactgatgg gtcttggtct
     4141 cttgcatctg agccttccca gcaggacctg ttctcagccg cagctcctta caactttggg
     4201 cacttcaacg actccgaaat taccaaggat ctgaacgata ttgattctgc caaatctgag
     4261 aaccctactt accggaaggc cgcttttgta aagtatcagg aagatatgaa taagaaggca
     4321 tacgtgatcc caactaactt tatgctgaac tacactccag tgaataagcg cgtggttggc
     4381 atgaccctgg attacggagc tatgaatacc tggagtgaga ttggtgtcag tagcgctaag
     4441 ctggcaacaa agtgagaatt cgaagcttga tccggctgct aacaaagccc gaaaggaagc
     4501 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     4561 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatctggcg taatagcgaa
     4621 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatg
//
