LOCUS       Exported                9528 bp ds-DNA     circular SYN 23-JUL-2020
DEFINITION  synthetic circular DNA
ACCESSION   MesoLSHC416B
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9528)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jul 23, 2020 from SnapGene Viewer 5.1.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9528
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          52..4844
                     /mol_type="other DNA"
                     /label=for use in Methylobacterium extorquens AM1
                     /label=for use in Methylobacterium extorquens AM1; 
                     theHindIII-BamHI fragment of pCM27 (pCR2.1 with a PCR f
                     /note="for use in Methylobacterium extorquens AM1; 
                     theHindIII-BamHI fragment of pCM27 (pCR2.1 with"
                     /db_xref="taxon:163915"
     source          4845..7296
                     /label=for use in Methylobacterium extorquensAM1;
                     the ...
                     /label=source
     protein_bind    1..24
                     /label=TEV
     misc_binding    25..48
                     /label=His-tag
     gene            57..380
                     /gene="lacZa"
                     /label=lacZa
     CDS             57..380
                     /codon_start=1
                     /transl_table=11
                     /gene="lacZa"
                     /product="LacZ"
                     /label=LacZ
                     /note="beta-galactosidase alpha peptide"
                     /protein_id="AAK73405.1"
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     CDS             complement(568..1233)
                     /codon_start=1
                     /transl_table=11
                     /gene="tetR"
                     /product="TetR"
                     /function="tetracycline resistance regulation"
                     /label=TetR
                     /protein_id="AAK73406.1"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGAKKPLLS"
     gene            complement(568..1233)
                     /gene="tetR"
                     /label=tetR
     gene            1339..2538
                     /gene="tetA"
                     /label=tetA
     CDS             1339..2538
                     /codon_start=1
                     /transl_table=11
                     /gene="tetA"
                     /product="TetA"
                     /function="tetracycline resistance"
                     /label=TetA
                     /protein_id="AAK73407.1"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     gene            complement(2769..3917)
                     /gene="trfA"
                     /label=trfA
     CDS             complement(2769..3917)
                     /codon_start=1
                     /transl_table=11
                     /gene="trfA"
                     /product="TrfA"
                     /function="trans-acting replication"
                     /label=TrfA
                     /protein_id="AAK73408.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     oriT            4099..4837
                     /label=IncP origin of transfer
                     /note="IncP origin of transfer"
     CDS             complement(4444..4560)
                     /codon_start=1
                     /transl_table=11
                     /gene="traJ'"
                     /product="TraJ'"
                     /label=TraJ'
                     /note="truncated oriT-recognizing protein"
                     /protein_id="AAK73409.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL"
     gene            complement(4444..4560)
                     /gene="traJ'"
                     /label=traJ'
     rep_origin      4848..5556
                     /label=oriV
                     /label=oriV; IncP origin of replication
                     /note="oriV; IncP origin of replication"
     rep_origin      6024..6579
                     /label=colE1
                     /note="colE1"
     promoter        6861..6916
                     /label=Plac
                     /label=Plac; lac operon promoter/operator
                     /note="Plac; lac operon promoter/operator"
                     /note="/regulatory_class=promoter"
     promoter        6962..7279
                     /label=PmxaF
                     /label=PmxaF; Methylobacterium extorquens mxaoperonpromoter
                     /note="PmxaF; Methylobacterium extorquens 
                     mxaoperonpromoter"
                     /note="/regulatory_class=promoter"
     promoter        7301..7612
                     /label=PxoxF, xoxF promoter
     misc_feature    7630..7646
                     /label=T7_transl_en_RBS
     misc_feature    7654..9528
                     /label=MesoLSHC416B
ORIGIN
        1 gccgaaaacc tctattttca gggacatcat caccatcacc atcaccactg acagctatga
       61 ccatgattac gccaagcttg catgcctgca ggtcgactct agaggatccc cgggtaccga
      121 gctcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
      181 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
      241 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
      301 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
      361 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
      421 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
      481 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
      541 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag caccctttct
      601 cggtccttca acgttcctga caacgagcct ccttttcgcc aatccatcga caatcaccgc
      661 gagtccctgc tcgaacgctg cgtccggacc ggcttcgtcg aaggcgtcta tcgcggcccg
      721 caacagcggc gagagcggag cctgttcaac ggtgccgccg cgctcgccgg catcgctgtc
      781 gccggcctgc tcctcaagca cggccccaac agtgaagtag ctgattgtca tcagcgcatt
      841 gacggcgtcc ccggccgaaa aacccgcctc gcagaggaag cgaagctgcg cgtcggccgt
      901 ttccatctgc ggtgcgcccg gtcgcgtgcc ggcatggatg cgcgcgccat cgcggtaggc
      961 gagcagcgcc tgcctgaagc tgcgggcatt cccgatcaga aatgagcgcc agtcgtcgtc
     1021 ggctctcggc accgaatgcg tatgattctc cgccagcatg gcttcggcca gtgcgtcgag
     1081 cagcgcccgc ttgttcctga agtgccagta aagcgccggc tgctgaaccc ccaaccgttc
     1141 cgccagtttg cgtgtcgtca gaccgtctac gccgacctcg ttcaacaggt ccagggcggc
     1201 acggatcact gtattcggct gcaactttgt catgattgac actttatcac tgataaacat
     1261 aatatgtcca ccaacttatc agtgataaag aatccgcgcg ttcaatcgga ccagcggagg
     1321 ctggtccgga ggccagacgt gaaacccaac atacccctga tcgtaattct gagcactgtc
     1381 gcgctcgacg ctgtcggcat cggcctgatt atgccggtgc tgccgggcct cctgcgcgat
     1441 ctggttcact cgaacgacgt caccgcccac tatggcattc tgctggcgct gtatgcgttg
     1501 gtgcaatttg cctgcgcacc tgtgctgggc gcgctgtcgg atcgtttcgg gcggcggcca
     1561 atcttgctcg tctcgctggc cggcgccact gtcgactacg ccatcatggc gacagcgcct
     1621 ttcctttggg ttctctatat cgggcggatc gtggccggca tcaccggggc gactggggcg
     1681 gtagccggcg cttatattgc cgatatcact gatggcgatg agcgcgcgcg gcacttcggc
     1741 ttcatgagcg cctgtttcgg gttcgggatg gtcgcgggac ctgtgctcgg tgggctgatg
     1801 ggcggtttct ccccccacgc tccgttcttc gccgcggcag ccttgaacgg cctcaatttc
     1861 ctgacgggct gtttcctttt gccggagtcg cacaaaggcg aacgccggcc gttacgccgg
     1921 gaggctctca acccgctcgc ttcgttccgg tgggcccggg gcatgaccgt cgtcgccgcc
     1981 ctgatggcgg tcttcttcat catgcaactt gtcggacagg tgccggccgc gctttgggtc
     2041 attttcggcg aggatcgctt tcactgggac gcgaccacga tcggcatttc gcttgccgca
     2101 tttggcattc tgcattcact cgcccaggca atgatcaccg gccctgtagc cgcccggctc
     2161 ggcgaaaggc gggcactcat gctcggaatg attgccgacg gcacaggcta catcctgctt
     2221 gccttcgcga cacggggatg gatggcgttc ccgatcatgg tcctgcttgc ttcgggtggc
     2281 atcggaatgc cggcgctgca agcaatgttg tccaggcagg tggatgagga acgtcagggg
     2341 cagctgcaag gctcactggc ggcgctcacc agcctgacct cgatcgtcgg acccctcctc
     2401 ttcacggcga tctatgcggc ttctataaca acgtggaacg ggtgggcatg gattgcaggc
     2461 gctgccctct acttgctctg cctgccggcg ctgcgtcgcg ggctttggag cggcgcaggg
     2521 caacgagccg atcgctgatc gtggaaacga taggcctatg ccatgcgggt caaggcgact
     2581 tccggcaagc tatacgcgcc ctagaattgt caattttaat cctctgttta tcggcagttc
     2641 gtagagcgcg ccgtgcgtcc cgagcgatac tgagcgaagc aagtgcgtcg agcagtgccc
     2701 gcttgttcct gaaatgccag taaagcgctg gctgctgaac ccccagccgg aactgacccc
     2761 acaaggccct agcgtttgca atgcaccagg tcatcattga cccaggcgtg ttccaccagg
     2821 ccgctgcctc gcaactcttc gcaggcttcg ccgacctgct cgcgccactt cttcacgcgg
     2881 gtggaatccg atccgcacat gaggcggaag gtttccagct tgagcgggta cggctcccgg
     2941 tgcgagctga aatagtcgaa catccgtcgg gccgtcggcg acagcttgcg gtacttctcc
     3001 catatgaatt tcgtgtagtg gtcgccagca aacagcacga cgatttcctc gtcgatcagg
     3061 acctggcaac gggacgtttt cttgccacgg tccaggacgc ggaagcggtg cagcagcgac
     3121 accgattcca ggtgcccaac gcggtcggac gtgaagccca tcgccgtcgc ctgtaggcgc
     3181 gacaggcatt cctcggcctt cgtgtaatac cggccattga tcgaccagcc caggtcctgg
     3241 caaagctcgt agaacgtgaa ggtgatcggc tcgccgatag gggtgcgctt cgcgtactcc
     3301 aacacctgct gccacaccag ttcgtcatcg tcggcccgca gctcgacgcc ggtgtaggtg
     3361 atcttcacgt ccttgttgac gtggaaaatg accttgtttt gcagcgcctc gcgcgggatt
     3421 ttcttgttgc gcgtggtgaa cagggcagag cgggccgtgt cgtttggcat cgctcgcatc
     3481 gtgtccggcc acggcgcaat atcgaacaag gaaagctgca tttccttgat ctgctgcttc
     3541 gtgtgtttca gcaacgcggc ctgcttggcc tcgctgacct gttttgccag gtcctcgccg
     3601 gcggtttttc gcttcttggt cgtcatagtt cctcgcgtgt cgatggtcat cgacttcgcc
     3661 aaacctgccg cctcctgttc gagacgacgc gaacgctcca cggcggccga tggcgcgggc
     3721 agggcagggg gagccagttg cacgctgtcg cgctcgatct tggccgtagc ttgctggacc
     3781 atcgagccga cggactggaa ggtttcgcgg ggcgcacgca tgacggtgcg gcttgcgatg
     3841 gtttcggcat cctcggcgga aaaccccgcg tcgatcagtt cttgcctgta tgccttccgg
     3901 tcaaacgtcc gattcattca ccctccttgc gggattgccc cgactcacgc cggggcaatg
     3961 tgcccttatt cctgatttga cccgcctggt gccttggtgt ccagataatc caccttatcg
     4021 gcaatgaagt cggtcccgta gaccgtctgg ccgtccttct cgtacttggt attccgaatc
     4081 ttgccctgca cgaataccag ctccgcgaag tcgctcttct tgatggagcg catggggacg
     4141 tgcttggcaa tcacgcgcac cccccggccg ttttagcggc taaaaaagtc atggctctgc
     4201 cctcgggcgg accacgccca tcatgacctt gccaagctcg tcctgcttct cttcgatctt
     4261 cgccagcagg gcgaggatcg tggcatcacc gaaccgcgcc gtgcgcgggt cgtcggtgag
     4321 ccagagtttc agcaggccgc ccaggcggcc caggtcgcca ttgatgcggg ccagctcgcg
     4381 gacgtgctca tagtccacga cgcccgtgat tttgtagccc tggccgacgg ccagcaggta
     4441 ggcctacagg ctcatgccgg ccgccgccgc cttttcctca atcgctcttc gttcgtctgg
     4501 aaggcagtac accttgatag gtgggctgcc cttcctggtt ggcttggttt catcagccat
     4561 ccgcttgccc tcatctgtta cgccggcggt agccggccag cctcgcagag caggattccc
     4621 gttgagcacc gccaggtgcg aataagggac agtgaagaag gaacacccgc tcgcgggtgg
     4681 gcctacttca cctatcctgc ccggctgacg ccgttggata caccaaggaa agtctacacg
     4741 aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat ggatataccg aaaaaatcgc
     4801 tataatgacc ccgaagcagg gttatgcagc ggaaaagatc cgtcgaccct ttccgacgct
     4861 caccgggctg gttgccctcg ccgctgggct ggcggccgtc tatggccctg caaacgcgcc
     4921 agaaacgccg tcgaagccgt gtgcgagaca ccgcggccgc cggcgttgtg gatacctcgc
     4981 ggaaaacttg gccctcactg acagatgagg ggcggacgtt gacacttgag gggccgactc
     5041 acccggcgcg gcgttgacag atgaggggca ggctcgattt cggccggcga cgtggagctg
     5101 gccagcctcg caaatcggcg aaaacgcctg attttacgcg agtttcccac agatgatgtg
     5161 gacaagcctg gggataagtg ccctgcggta ttgacacttg aggggcgcga ctactgacag
     5221 atgaggggcg cgatccttga cacttgaggg gcagagtgct gacagatgag gggcgcacct
     5281 attgacattt gaggggctgt ccacaggcag aaaatccagc atttgcaagg gtttccgccc
     5341 gtttttcggc caccgctaac ctgtctttta acctgctttt aaaccaatat ttataaacct
     5401 tgtttttaac cagggctgcg ccctgtgcgc gtgaccgcgc acgccgaagg ggggtgcccc
     5461 cccttctcga accctcccgg cccgctaacg cgggcctccc atccccccag gggctgcgcc
     5521 cctcggccgc gaacggcctc accccaaaaa tggcagccaa gctgaccact tctgcgctcg
     5581 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5641 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5701 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5761 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5821 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5881 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5941 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6001 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6061 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     6121 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6181 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6241 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6301 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6361 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6421 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6481 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6541 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6601 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     6661 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     6721 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     6781 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     6841 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     6901 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctagct
     6961 tcccgcttgg tcgggccgct tcgcgagggc ccgttgacga caacggtgcg atgggtcccg
     7021 gccccggtca agacgatgcc aatacgttgc gacactacgc cttggcactt ttagaattgc
     7081 cttatcgtcc tgataagaaa tgtccgacca gctaaagaca tcgcgtccaa tcaaagccta
     7141 gaaaatatag gcgaagggac gctaataagt ctttcataag accgcgcaaa tctaaaaata
     7201 tccttagatt cacgatgcgg cacttcggat gacttccgag cgagcctgga acctcagaaa
     7261 aacgtctgag agataccgcg gatctcacac aggaaatttc gcaacgcaaa atgctgaatt
     7321 tagcaggcaa gtttcctgat aaagggccct gtgagtgcga cattcgacgc gcgaaagcga
     7381 gttgacggcg cccgaaattt tcttagagtt cgttcaagct tcggtttcgg ggagccgcta
     7441 agggatgcgg acccagccgg gtaagtcgcc gggaactatg gaataatcct aatttctgtg
     7501 cggcaggcgc gagctgccgg agcgaatatc atccctgtcg caaacagcag gtcaatcaac
     7561 ggtgggtagc gggctgctat ccagcggcgg ataagacttg tcggaggaat cctctagaaa
     7621 taattttgtt taactttaag aaggagatat accatgaagg cgatgccgca gctcgcgctg
     7681 acccgcgtcg acgtggagcg ccgtacacgc aagctggccg ccggaaacag cttcggcgcg
     7741 atcctcctgg tcaccctcct cgcgtgcttg ctgctcttgg gcgccgccag cgatccgccg
     7801 aagacgccgg cgatcgtgcc gccggccgcg gcgtccccgc cggacgatgg ccagtggacc
     7861 atgcccgcca agaactatgc ctccacccgc ttctcggagc tctccgagat caacgaggga
     7921 aacgtgaaga acctccaggt cgcgttcacc ttctccacgg gcgtgaacaa gggccaggag
     7981 gccgcgcctc tcgtggtcgg caacacgatg tacatcgtca ccccgttccc gaacttcgtc
     8041 tacgccctcg atctctcgaa gcccggtgcc cccatgaagt ggaagtacga gcccaacccc
     8101 gaacccgccg cgcagggcgt ggcgtgctgc gacgtggtca accgcggcgt tgccttcgcc
     8161 gatggccgca tcttcttcaa cacgctggac ggccacacca tcgccctgga cgcgaatacg
     8221 ggcaagccca tctggaacac gcacgtgggc aacatcaaca tcggcgagac catcaccatg
     8281 gcgccgctgg ttgtcaaggg gaaggtgctc gtcggcaact ccggcggcga gatgggggtc
     8341 cgcggctggg tgaaggcgct ggacgctggc gatggccatg tagtgtggat cgcgttctcg
     8401 accgggcccg acaaggacgt gctcatcggc cccgatttca agccgcatta cgacatggac
     8461 aagggcaagg acctcggggt gaccacctgg ccgcctgaag cctggaagat cggcggcggt
     8521 aacatgtggg gctggatctc ctacgatccc gatctcaacc tgatcttcca cggcaccggc
     8581 aaccccggcc cctggaaccc cgacctgcgc ccgggcgaca acaagtggac ctccggcatc
     8641 ttcgcgcggg acccagatac cggtgccgcg aagtggttct accagtggag cccgcacgac
     8701 ctccacgact acgacggcat caacgagcag atcctcctgg acatgacctg gcagggcaag
     8761 ccgcggaagg tcctgatccg gccggaacgc aacggctacc tgtacatcct ggaccgcacc
     8821 accggcgagg tgctgtcggc caagccgttc ggccctgtca actcctcgaa gggggtggat
     8881 ctgaagaccg gccgcctcat cgtgaacccg gacaagaaga cggggaccgg caaggtggtc
     8941 cgggacatct gccccacggc gtcgggactg aaggactggc agccgtcggc cttctcgccg
     9001 aagaccggcc tgctgtacat cccgcacaac aacctgtgca tggacgagga gggtgtcgag
     9061 gtcaactata tcgctggcac cccgtacgtc ggaatgaacg tccgcatgat cccgggcccc
     9121 ggcggcaacc gcggggcgtt caccgcctgg gacatcgcag cggagaagcc ggcctggagc
     9181 ctcaaggaaa attttccagt gtggtcggga gcagtggtga ccgcgggcga tgtcgtgttc
     9241 tacggcacca tggaaggctg gttcaaggcg gtctcggcga agacgggcga cctgctctgg
     9301 cagtttaaga ccagcagcgg catcatcggg cagcccatca cttaccgcgg cccggatggg
     9361 caccagtatg tcgcgatcct gtcgggcgtt ggcggctggg cgggcgcgat cgtgagcgga
     9421 gacctcgacc cccgcgacgc cacggccgcg ctgggcttcg tgaacgcgat gaaggacttg
     9481 aagaatgcga ccacctcggg cggcaccttg tacgttttcc ggctcccc
//
