LOCUS       pMVS1111A-PhmtB_       10403 bp DNA     circular SYN 25-SEP-2023
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10403)
  AUTHORS   Lucas Muhling
  TITLE     Direct Submission
  JOURNAL   Exported Sep 25, 2023 from SnapGene 7.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(82..705)
                     /gene="catP"
                     /product="Chloramphenicol reistance for E. coli"
                     /label=catP
     CDS             879..1673
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     terminator      1688..2032
                     /label=Tmcr
     misc_feature    complement(2095..4113)
                     /label=beta galactosidase codon opt M.t
                     /label=beta galactosidase codon opt M.t.
                     /note="used for black white screening "
     promoter        complement(4114..4206)
                     /label=PhmtB
     misc_feature    4253..8642
                     /label=pME2001 (Methanothermobacter ori)
     terminator      8690..8743
                     /gene="CD0164 terminator"
                     /label=CD0164 terminator
     CDS             complement(8886..9326)
                     /gene="traJ"
                     /label=traJ
     oriT            9290..9399
                     /label=incP origin of transfer
                     /note="incP origin of transfer"
     rep_origin      complement(9720..10308)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aaacttaggg taacaaaaaa caccgtattt ctacgatgtt tttgcttaaa tacttgtttt
       61 tagttacaga caaacctgaa gttaactatt tatcaattcc tgcaattcgt ttacaaaacg
      121 gcaaatgtga aatccgtcac atactgcgtg atgaacttga attgccaaag gaagtataat
      181 tttgttatct tctttataat atttccccat agtaaaaata ggaatcaaat aatcatatcc
      241 tttctgcaaa ttcagattaa agccatcgaa ggttgaccac ggtatcatag atacattaaa
      301 aatgttttcc ggagcatttg gctttccttc cattctatga ttgtttccat accgttgcgt
      361 atcactttca taatctgcta aaaatgattt aaagtcagac ttacactcag tccaaaggct
      421 ggaaaatgtt tcagtatcat tgtgaaatat tgtatagctt ggtatcatct catcatatat
      481 ccccaattca ccatcttgat tgattgccgt cctaaactct gaatggcggt ttacaatcat
      541 tgcaatataa taaagcattg caggatatag tttcattccc ttttccttta tttgtgtgat
      601 atccacttta acggtcatgc tgtaggtaca aggtacactt gcaaagtagt ggtcaaaata
      661 ctcttttctg ttccaactat ttttatcaat tttttcaaat accatctaag ttccctctca
      721 aattcaagtt tatcgctcta atgaacaaag atattatacc acatttttgt gaatttttca
      781 acttgcccac tggccggccg cccatttttt gagtattctt tatataggga tatagtaata
      841 gataatatca cggtaaccac cggcagcagg tgatgcatat ggctaaaatg agaatatcac
      901 cggaattgaa aaaactgatc gaaaaatacc gctgcgtaaa agatacggaa ggaatgtctc
      961 ctgctaaggt atataagctg gtgggagaaa atgaaaacct atatttaaaa atgacggaca
     1021 gccggtataa agggaccacc tatgatgtgg aacgggaaaa ggacatgatg ctatggctgg
     1081 aaggaaagct gcctgttcca aaggtcctgc actttgaacg gcatgatggc tggagcaatc
     1141 tgctcatgag tgaggccgat ggcgtccttt gctcggaaga gtatgaagat gaacaaagcc
     1201 ctgaaaagat tatcgagctg tatgcggagt gcatcaggct ctttcactcc atcgacatat
     1261 cggattgtcc ctatacgaat agcttagaca gccgcttagc cgaattggat tacttactga
     1321 ataacgatct ggccgatgtg gattgcgaaa actgggaaga agacactcca tttaaagatc
     1381 cgcgcgagct gtatgatttt ttaaagacgg aaaagcccga agaggaactt gtcttttccc
     1441 acggcgacct gggagacagc aacatctttg tgaaagatgg caaagtaagt ggctttattg
     1501 atcttgggag aagcggcagg gcggacaagt ggtatgacat tgccttctgc gtccggtcga
     1561 tcagggagga tatcggggaa gaacagtatg tcgagctatt ttttgactta ctggggatca
     1621 agcctgattg ggagaaaata aaatattata ttttactgga tgaattgttt tagcgcccgc
     1681 cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatggctcc gaccgaagcc
     1741 acccggggcg gccccgccga ccccgcaccc gcccccgagg cccaccgcgg gggacacacc
     1801 gaacacgccg accctgctga acacgcggcg cagttcggtg cccaggagcg gatcgggaat
     1861 taattcgaag ctgctggtga aagagaccct atcttacctg ctaaaatcta agttaattac
     1921 taatttatta ttaatttatt attagattgg gcaaaatagt aaaagaaaac taaaggaaac
     1981 ctaatatggt ttcctttttt tatatatttt taattcactg ggggcaattc tgtcagggcg
     2041 cgcctggggt cgtgcgctcc tttcggtcgg gcgctgcggg tcgtggggcg ggcgctagac
     2101 cttgccggct tcgtcgtgtt ccctaaggac agcgacgtcg acgccctgaa tcctgagttc
     2161 accccccctg aagcatttgc catctatcat attctggtag atcttatctt ccggaaggga
     2221 gagtgtgacc tcatagtcgt tgtggttaat tataataagg tacttccatt catcggtctc
     2281 cctctgctga acttcgacat tctcagcaac ctccagtata ggatttatgt ggtgtttagc
     2341 aaacacctgt tcgagaagcc tgccaaggta gttgctgtca gggtatgttc ctacgtatat
     2401 gccctccccc tttccgtagc agttcctggt aacagcagga aggccggcat accaatcacc
     2461 tttgaatgtg gcgagaggct cagcaccttc cagccttatt atatcggccc atgtggtaca
     2521 gtcatactcg ccgtcgtttg agtagatctt attcaccttt gtctcgggat aaggaacgaa
     2581 ttcctccacg aagatgccga gaatgtccct cagcggtcct ggatatcccc cgaggtgcac
     2641 tctatcgttc tcatcgacta tcacactgaa aaagcttaca atcagggttc cgccgtttgc
     2701 gacaaactgc ctaaggttct catcttctcc ctctttcacc atatacagca tcggtgcaat
     2761 aacaacctta tattttgtga gatcgtcgga cggtcttaca aagtcgactg ctatgtttct
     2821 cttgtaaagc tctctataat atgcctctac tatgggaata tatctgagct tgttgtgcgg
     2881 tttggaactg agctcaactg cccaccagtt ttcccagtca aagataattg ccacctctgc
     2941 ctttattcta ctccccacga ggcagtcaag ttttttcagc tcctggccaa gctgggtaac
     3001 ttccctgtat attctattgt tttcgttaag aaagtggggc accattgctc cgtgaaactt
     3061 ctcagctcct gctctggact gcctccactg aaagaacatt atcccatcgg cacccctggc
     3121 gattgttgcg taactccaga gtctcataac ccccggcggc tttggcacat tgatatctct
     3181 ccaattaacg tgactggtga cctgctccat aagaatgaac ggctgcccct tcctaagtga
     3241 cctcatgagg tcattcatca ttgcgtgctg tatagggagt ccctccctgg gatctgggta
     3301 gctatcccag gtaacgatat ctacgtgctg agcccactga aagtagttga gtggcttgaa
     3361 tgatcccatg aaatttgtgg agaccgggat atcgggggtt acttccctga ggatctcctt
     3421 ttctgtaagg aagagtttga ggattgaatc attcatgaat ctgtagtaat caagctcctg
     3481 gctggggtta ataaatgttg gtgccttcct agggggatta atctcatccc agtggttata
     3541 tctctggccc cagaagtttg taccccatct ttcattaagt tcatcaatgg tcttatacct
     3601 ttctttaagc cattttctga aagcaactgc gcaattctca cagaaacact tacttacatg
     3661 gcaagcgtat tcgttattta cgtgccacat tttgagggct ggatgatttt tgtatctctc
     3721 agctatagcc cttaccagcc tctttatatg tgttataagc tgagggtgat ttgggcaata
     3781 atgctgtcta ctcccgaaac tcagtatcac accggactcg tcaataggga gtgaatcagg
     3841 gtatttcttc acgaaccagg cgggtgtggt tgcggtggcg gtccccagat ttatgtatac
     3901 cccatgatcg tagaggatgt ctatcacttt gtcgagccat tcaaaatcaa atacaccgtc
     3961 tgatggctcg attttggacc agctaaagat tccgagtgaa acaagattaa caccggcctt
     4021 ctgcataagt tttgcgtcct cgtaccatat ctcctcgggc cactgttctg ggttgtaatc
     4081 ccccccatag cagatggaac tgagaacgtt cataattaac cacctcattt tatgtgatat
     4141 tatataatcc tatataaata tatcgctaat tttaaggttt ttctgagcca tcggttggtt
     4201 catgggggca ttcctccagg cttaattaac aataaaattt ggggtggcag ggtggagtaa
     4261 aagtgtagtg tgatgatagt ggattccatc tacctggggt gtcatccccc actatggggt
     4321 tgtgtttttc tgtggttttg tgtgtgtttc tgtggttgta gtgacacttt acttcggtat
     4381 atcattatat tacgaagtca aatcatggta ctgtgccgat gttatggggt taattgttta
     4441 tactgtgaca caccatagtt atatgatatg atcataggag tggattgtat ggcgatggag
     4501 gatgttggcg tgccattcag taacaggtta accaggcagg ggaacattaa ggtgcctgct
     4561 gacttgaggg atgctttgaa gttgaagcct ggtgacctcc tggtggtgga gatcaaaaaa
     4621 gttgacaggt catagtgaca ctctccccta ttggcgaggc attttaactc tttttatgct
     4681 tttacggttt tgcagaaaat cgcattccct gtaagataat ataagaaaaa ataatatagt
     4741 atattaactg ttttcatggc ccgggctaag ttttaatagc aagaacaaca agaaatagtt
     4801 atgaaaaagc gttgttgagt ggggtggaaa taccgcacac caaaacccca ccaacacagt
     4861 caccctgcag aaaatgtagg ggtacacacc ttgcagaatg gtaccctgca taagggttgt
     4921 aaagccctgg cttattctat gactgccagg gcatataaaa tcttccctcg attctgcacc
     4981 ttgattctgc agggtgacac caaaaacaaa aggggtgatc accttggata aaaaggcgat
     5041 aacagtcgag ataggtatag aggaatactt gaaactgcag gatttaaggg ttgaatccct
     5101 caagactggt aggtttgtgt ctgttgggtg tgtcatgagg gatgccctta aacactacct
     5161 gcaggaggtt gacctgagag agaaatacct ccagcaggca gaggacaata agaaggagtt
     5221 gataagatga gtggcaacca caaccaccca ccccacagaa ccacccctat agaggtgatg
     5281 tgcaatccac ccacaaagga ggtgaactac aatgaatagt atgtcaaaga aggaatataa
     5341 acattttgag cgatcaataa ccctacacat taaacacgct gattttgaat tagtaggcca
     5401 gaaatacagg ggcagagacc tctacgagta cttgttcatc aaggggagta aaccaatcca
     5461 cacagccaca ggtaaaacca gcaacctcta cgagatcata tcagaggata aaggccctga
     5521 tgaagccctc aaaatcatag gggacacatt cacagaggat gatataaaca tcctcctcag
     5581 gggagggttc catgatgata acaagacccc tgagggtgtc ctcgagttaa tccagcacat
     5641 cctcctcgca ggtgaggtgc tccacccagg aggggatgtt attgaacccc aatcattcaa
     5701 ggactaccct gagaagatac aagcctacgc tgaccagtta atcaatgatg acagcatcga
     5761 catcctcgaa tccatcacca gagtcatagg ggaggcccat tatggtgatg agaaggccgt
     5821 gaaattactc ctattatcca tcggcaccct attcctcagg gacaccccac cagtccacca
     5881 ggcactgagg ggctccacag gttcaggtaa aacagacctt gtattgaaga cagtactcgc
     5941 agtcccagag aggtatgtgc acatcctcag atcagcatca cctaaatact tattctacgc
     6001 atcagagact ggcatcctcc gtgaagacta caatatcttc gtatttgatg atattgagtt
     6061 gaatgatgag atcatagcga tctccaagac catcacagat aacatcctcc cagagaagga
     6121 acaccacacc gtgaaggatc aggaggccct gaaactggag atcccaggtg agggcctggc
     6181 gatattcaca agggcgaggg acatccatga taatgaactc aatgacaggc tactctacaa
     6241 taacccagtg gaggatgagg atcacagcag atttgtgaag gagaagatca aagaggaggc
     6301 catctcaggg tcagtgatgg atgataagag gatcatggag gcctacgagg tcatcagggc
     6361 tgtctatgag cgattgatag agggtgatgt cagggtttac aacccctacc tccacctact
     6421 cgacctgaga gggtatagta acagggacat caagcacata gttggacttg tgaaggccgt
     6481 gacatggtac aggcaacata aaaggaggag ggaggggagt tatgtccgtg aaggtggcca
     6541 tgacattatc atcggcactg aggatgacct gagggatgca ttggagttgt ggatgtccat
     6601 agacaccctc caaaggtata agttggacag gaaacaggag aaattcctca aatccctacc
     6661 agaatactct gatgagttat ataaacagca tactaattca atcttttggg atgatcctga
     6721 acttccaact tatagtggtc tcgctgagag gcttgggttg agtaaatcaa caatatatcg
     6781 gtgggttaat gggaggcatg agaaagacag agacatcgag ggcttaaagg ataaggggaa
     6841 taagggtctt gtcattatca aaccactgga acctgagaat cctaaatcac cctcattgat
     6901 ataccgcaaa ccaggtattg ttgggaaaca cctccaactc aaaaatggcg gagaagcctc
     6961 agaatctgga tttaccctgt tccccagtgt tgacatggac ttgggaacag cggaaaatgc
     7021 tcggatttgg aaaaaaacca tattccatag tatggtttta ggtgtttccc aaattcaaag
     7081 taacatacaa gacaatttaa tacaagaatg gcttgataat aatactaatc ccatggagac
     7141 acatgaggat gttgttgaat tcattgtcga tgtcagggcc ttcatccatg acaacctgca
     7201 gacacctcct gatgtgacct gcaacccagt atataccata ctgggaaatg ggaaacaccc
     7261 cctggaaaca ggtggagaat cctccaatca cggagataag catgatttgg tgttgaccag
     7321 ggccgggtca atagtgggga atagggtaaa cagtgaaaat aagggctctg agccctccga
     7381 tgaaatgaca tgtttccccc aaaaaaatta ttactctggt gtgggtggtg atctgcaggc
     7441 tgagtacact gatgcaatgg cacggggtga tgactcgatg ataggtgaac ttgaggcgag
     7501 gatcccacca gacacacctg agaaccttga ggagaggagg gtcaccctga aaaagaaact
     7561 caaggataag agtatcagtg ccggggagaa gatggaactc ctcaaggaac tgggaggggt
     7621 gctctcaact ctaatgagga aaccagtgga taatgagagg attgtcgagt atgcgaagga
     7681 attgggtgag gttcggcggg agatagagga tatgcagggg gggtgatcca ttatgcccct
     7741 ctccttagca acttcaaacc gtgtccctga ggcttggagg gatctgatca tatcggctat
     7801 gagtaacctc gcacccctcg ggagggatcc gaatacaaag gaattcacag atgaactcca
     7861 ccacctctcg ggggaagata tggggactgt gatggaaatc ctggggattg cagtggctga
     7921 ggggttcctg gaatacgatg atcattacag gctcatcctc ggccctgcag gtgaggagta
     7981 tgttcggctg tcatcacggg ttaaggttga ggagacaagg agattgacac ttcgaagggc
     8041 cctcaggaga ttcagggata aaaggagggc ctgcaggcac ctgtcccttg gggataaggc
     8101 cctttatgat tactattacc atggtggcct gtgaattata attcaatgtc cttgttgggt
     8161 ggaatgtgga agtcaacaaa accctcaaga tccaaatcac caaagccttc aagtaacccc
     8221 ttaaggtctt caggtttgaa ctctgggatg tcatcaaggt taaactctaa gagttcctta
     8281 aggtcggggc attcaaggtc atcaaatact gggaggtctt caaggcaaaa gtctattagt
     8341 ggttcattag ggtcaaatcc tattagttct tccaggttga actctaaaac cctgttatca
     8401 tcaatcttca tgtgtaaata tctatatcct ccatggatta ataattttgg gaattttggg
     8461 agggttgaac caccaccctt gatagaaaaa aatctaaaaa aatctaaaaa aatcaaaata
     8521 aaaattcacc cctggactgg gttcccctat aatcaagtcc tgtctgagaa agcaaaaaaa
     8581 tacaagaaaa tccgtggatg actccattat gaatctattt tgaatccatt gcgttgcgct
     8641 gcgatcgcgg tcatatggat acagcggccg cggtcatagc tgtttcctga taaaaaaatt
     8701 gtagataaaa ctattttata aaatttatct acaatttttt tatcctgcag ggggcccgat
     8761 cggtcttgcc ttgctcgtcg gtgatgtact tcaccagctc cgcgaagtcg ctcttcttga
     8821 tggagcgcat ggggacgtgc ttggcaatca cgcgcacccc ccggccgttt tagcggctaa
     8881 aaaagtcatg gctctgccct cgggcggacc acgcccatca tgaccttgcc aagctcgtcc
     8941 tgcttctctt cgatcttcgc cagcagggcg aggatcgtgg catcaccgaa ccgcgccgtg
     9001 cgcgggtcgt cggtgagcca gagtttcagc aggccgccca ggcggcccag gtcgccattg
     9061 atgcgggcca gctcgcggac gtgctcatag tccacgacgc ccgtgatttt gtagccctgg
     9121 ccgacggcca gcaggtaggc cgacaggctc atgccggccg ccgccgcctt ttcctcaatc
     9181 gctcttcgtt cgtctggaag gcagtacacc ttgataggtg ggctgccctt cctggttggc
     9241 ttggtttcat cagccatccg cttgccctca tctgttacgc cggcggtagc cggccagcct
     9301 cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa
     9361 cacccgctcg cgggtgggcc tacttcacct atcctgcccg gctgacgccg ttggatacac
     9421 caaggaaagt ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga
     9481 tataccgaaa aaatcgctat aatgaccccg aagcaggggg ccctgcgtat tgggcgctct
     9541 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     9601 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     9661 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     9721 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     9781 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     9841 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     9901 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     9961 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
    10021 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
    10081 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
    10141 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
    10201 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
    10261 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
    10321 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
    10381 atgagattat caaaaaggag ttt
//
