LOCUS       V015867                 7711 bp    DNA     circular SYN 10-MAR-2019
DEFINITION  Exported.
ACCESSION   V015867
VERSION     V015867
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7711)
  AUTHORS   Cui J.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-JUN-2018) MCB, Dartmouth College, 14 Engineering
            Drive, Hanover, NH 03755, USA
REFERENCE   2  (bases 1 to 7711)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
            (28-JUN-2018) MCB, Dartmouth College, 14 Engineering Drive, Hanover,
            NH 03755, USA"
FEATURES             Location/Qualifiers
     source          1..7711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(100..1047)
                     /label="Rep101"
                     /note="RepA protein needed for replication with the pSC101
                     origin"
     rep_origin      complement(1095..1317)
                     /direction=LEFT
                     /label="pSC101 ori"
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     misc_feature    1844..2366
                     /label="upstream homology recombination flank"
                     /note="upstream homology recombination flank"
     gene            2367..3212
                     /gene="nfnA"
                     /label="nfnA"
     CDS             2367..3212
                     /codon_start=1
                     /transl_table=11
                     /gene="nfnA"
                     /product="NADH-dependent reduced ferredoxin:NADP
                     oxidoreductase subunit alpha"
                     /EC_number="1.6.1.4"
                     /label="nfnA"
                     /note="Clo1313_1848"
                     /protein_id="QBI89790.1"
                     /translation="MFKIVKKKILNPQVKLMEISAPHVAKKAEPGQFIILRVNENGERI
                     PLTIADFDREKGTVTIIFQEVGKTTKLLGTLEEGDELLDFVGPLGKASHFENVKKAAVI
                     GGGLGTAIAYPQAKKLHSMGVEVHTIAGFRNKDLIILEDEMRAVSSKLFITTDDGSNGN
                     KGFVSDVLKKLIEEGNKYDLVVAIGPLIMMKVISELTRPYGIKTIVSMNPVMIDGTGMC
                     GGCRVTVGGEIKFACVDGPDFDGHLVDFDEAMRRQAMYKKEESLALEKHNCKLGVEKNA
                     "
     gene            3205..4599
                     /gene="nfnB"
                     /label="nfnB"
     CDS             3205..4599
                     /codon_start=1
                     /transl_table=11
                     /gene="nfnB"
                     /product="NADH-dependent reduced ferredoxin:NADP
                     oxidoreductase subunit beta"
                     /EC_number="1.6.1.4"
                     /label="nfnB"
                     /note="Clo1313_1849"
                     /protein_id="QBI89791.1"
                     /translation="MPNMSPKKVPMPEQDPNVRIKNFLEVALGYTEQMAMEEAQRCLNC
                     KHKPCVSGCPVNVKIPEFVQLIAQGKFEKAYNKIRETNNLPAICGRVCPQENQCEKFCV
                     RGIKGEPVAIGRLERFAADWHMKNGTTSYEKPEKNGKRVAVIGSGPASLTCASDLAKLG
                     YEVTIFEAFHVPGGVLMYGIPEFRLPKKLVQEEIETIKQLGVEIKTNMVIGKVYSIDEL
                     KAEGYDAIFIGSGAGLPSFMKIPGENLNGVYSANEFLTRINLMKAYEFPNCDTPVKVGK
                     NVAVVGGGNVAMDAARSAKRLGAENVYIVYRRSEAEMPARLEEIHHAKEEGILFKFLTN
                     PTRILGTDDGWVKGMECIEMELGEPDESGRRRPVPKPGSEHVIDVETVIIAIGQTPNPL
                     IASTTPGLATQKWGGIIVDENTGATNIEGVYAGGDAVTGAATVILAMGAGKKAAKAIDE
                     YLKNKK"
     misc_feature    4600..5164
                     /label="downstream homology recombination flank"
                     /note="downstream homology recombination flank"
     regulatory      5169..5699
                     /label="kanamycin promoter"
                     /note="kanamycin promoter"
                     /regulatory_class="promoter"
     gene            5700..6461
                     /gene="htk"
                     /label="htk"
     CDS             5700..6461
                     /codon_start=1
                     /transl_table=11
                     /gene="htk"
                     /product="highly thermostable kanamycin
                     nucleotidyltransferase"
                     /EC_number="2.7.7.46"
                     /label="htk"
                     /protein_id="QBI89792.1"
                     /translation="MKGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGRQTD
                     GPYSDIEMMCVLSTEGVEFSYEWTTGEWKAEVNFYSEEILLDYASRVEPDWPLTHGRFF
                     SILPIYDPGGYFEKVYQTAKSVEAQKFHDAICALIVEELFEYAGKWRNIRVQGPTTFLP
                     SLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQPDLPPGYVQLCQLVMSGQLSDPEK
                     LLESLENFWNGVQEWAERHGYIVDVSKRIPF"
     CDS             6485..7063
                     /gene="tdk"
                     /label="Thymidine kinase"
                     /note="Thymidine kinase from Thermoanaerobacter
                     pseudethanolicus (strain ATCC 33223 / 39E). Accession#:
                     B0K7G9"
     misc_feature    7158..7711
                     /label="gene target internal homology recombination flank"
                     /note="gene target internal homology recombination flank"
ORIGIN
        1 gacagttttc cctttgatat gtaacggtga acagttgttc tacttttgtt tgttagtctt
       61 gatgcttcac tgatagatac aagagccata agaacctcag atccttccgt atttagccag
      121 tatgttctct agtgtggttc gttgtttttg cgtgagccat gagaacgaac cattgagatc
      181 atacttactt tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc
      241 agttaaagca tcgtgtagtg tttttcttag tccgttatgt aggtaggaat ctgatgtaat
      301 ggttgttggt attttgtcac cattcatttt tatctggttg ttctcaagtt cggttacgag
      361 atccatttgt ctatctagtt caacttggaa aatcaacgta tcagtcgggc ggcctcgctt
      421 atcaaccacc aatttcatat tgctgtaagt gtttaaatct ttacttattg gtttcaaaac
      481 ccattggtta agccttttaa actcatggta gttattttca agcattaaca tgaacttaaa
      541 ttcatcaagg ctaatctcta tatttgcctt gtgagttttc ttttgtgtta gttcttttaa
      601 taaccactca taaatcctca tagagtattt gttttcaaaa gacttaacat gttccagatt
      661 atattttatg aattttttta actggaaaag ataaggcaat atctcttcac taaaaactaa
      721 ttctaatttt tcgcttgaga acttggcata gtttgtccac tggaaaatct caaagccttt
      781 aaccaaagga ttcctgattt ccacagttct cgtcatcagc tctctggttg ctttagctaa
      841 tacaccataa gcattttccc tactgatgtt catcatctga gcgtattggt tataagtgaa
      901 cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca
      961 gcataaaatt agcttggttt catgctccgt taagtcatag cgactaatcg ctagttcatt
     1021 tgctttgaaa acaactaatt cagacataca tctcaattgg tctaggtgat tttaatcact
     1081 ataccaattg agatgggcta gtcaatgata attactagtc cttttccttt gagttgtggg
     1141 tatctgtaaa ttctgctaga cctttgctgg aaaacttgta aattctgcta gaccctctgt
     1201 aaattccgct agacctttgt gtgttttttt tgtttatatt caagtggtta taatttatag
     1261 aataaagaaa gaataaaaaa agataaaaag aatagatccc agccctgtgt ataactcact
     1321 actttagtca gttccgcagt attacaaaag gatgtcgcaa acgctgtttg ctcctctaca
     1381 aaacagacct taaaacccta aaggcttaag tagcaccctc gcaagctcgg gcaaatcgct
     1441 gaatattcct tttgtctccg accatcaggc acctgagtcg ctgtcttttt cgtgacattc
     1501 agttcgctgc gctcacggct ctggcagtga atgggggtaa atggcactac aggcgccttt
     1561 tatggattca tgcaaggaaa ctacccataa tacaagaaaa gcccgtcacg ggcttctcag
     1621 ggcgttttat ggcgggtctg cggagtggtg aatccgttag cgaggtgccg ccggcttcca
     1681 ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt
     1741 atagggcggc gcctacaatc catgccaacc cgttccatgt gctcgccgag gcggcataaa
     1801 tcgccgtgac gatcagcggt ccaatgatcg aagttaggct ggtgctcatg gtttagggct
     1861 tggtgcaata ttgccagcag ttataaaagc tatttatcca gctacagcag aagtattggc
     1921 tgatgtatat agtcctatag ttcctggttt aaaaggactg cctgttgagg cggagtatgt
     1981 agcagaaaaa gttcaagaat ggctttttag tgtggggtgc attcaaaagt tatctgattt
     2041 tgggtttacg aaagatgata ttcctaatct tgtaaaactt gctaaaacaa ctccttccct
     2101 tgatgggttg ttatctattg cacctgttga agctactgaa tctgtcattg aaaagattta
     2161 tttaaaatct ctttaacttt gataaataat tgtcttgttt tgttaaattt attacgatgt
     2221 tgttaagtga ttatgtaatt tgttgttttt gacgtaaaaa tatttgacat tagtcatata
     2281 aatatataat atatcattaa gtgaaaaatt atgtctggta taaagccaga cataatttat
     2341 tacgaaatct tggaggtgtg aaagcaatgt ttaaaatagt taaaaagaag attctcaacc
     2401 cacaggttaa gttgatggaa atatcggcgc cgcacgtagc taaaaaagca gagccgggac
     2461 agttcataat cctgagggta aatgaaaatg gtgaaagaat tcccctcaca atagccgatt
     2521 ttgaccgtga aaaaggtacg gtgacaataa tttttcagga agtgggaaaa accacaaaac
     2581 tactgggaac tttggaagaa ggagacgaat tgctggactt tgtcggacct ctgggtaaag
     2641 catcccactt tgagaacgta aaaaaagcgg ccgtcattgg aggaggcttg ggaacggcaa
     2701 tagcatatcc ccaggccaaa aaacttcact ccatgggtgt tgaagtccat acgatagcgg
     2761 gtttcagaaa caaagacctg ataattcttg aagatgaaat gagagccgta agcagcaaac
     2821 tttttataac caccgatgac ggttccaacg gcaataaagg tttcgtaagc gacgtattaa
     2881 agaaacttat tgaagaagga aacaaatatg atttggttgt tgccattggg ccgctaatta
     2941 tgatgaaagt tatcagcgag ctcacaagac cctacggcat taaaaccatt gtaagcatga
     3001 atccggttat gattgacggt accggcatgt gcggaggctg cagggttacc gttggcggtg
     3061 aaatcaagtt cgcatgtgtg gacggacctg attttgacgg gcatctggtt gattttgacg
     3121 aagcaatgag aaggcaggca atgtataaaa aagaagaatc cttggctttg gaaaaacaca
     3181 attgcaaatt gggggtggag aaaaatgcct aacatgtcac cgaaaaaagt tcccatgccg
     3241 gagcaggacc caaacgtcag aatcaaaaac tttttggagg ttgctttagg atataccgag
     3301 caaatggcaa tggaagaagc tcaaaggtgt cttaactgca agcacaaacc ttgtgtttcc
     3361 ggctgtcccg taaacgtaaa aattcctgag tttgtacagc ttatcgctca gggaaaattt
     3421 gagaaagcct acaataaaat aagagaaacc aacaaccttc cggcaatatg cggcagagtc
     3481 tgtccgcagg aaaaccagtg tgaaaagttc tgtgtaaggg gtataaaagg tgaacctgtt
     3541 gccataggaa ggcttgaaag atttgcggcg gactggcaca tgaaaaacgg caccacttct
     3601 tatgaaaagc ctgaaaaaaa cggcaaaagg gtggcagtaa taggttcggg acctgcaagc
     3661 cttacctgtg caagcgacct ggccaaactc ggctacgaag taacaatctt cgaagccttt
     3721 cacgtgcccg gcggagtgtt gatgtacggt attccggaat tcaggcttcc aaagaaactg
     3781 gttcaggagg aaattgaaac cataaagcag ctgggtgtgg aaattaaaac aaatatggtt
     3841 ataggaaagg tttattccat tgacgaactc aaagctgaag gatatgatgc catatttata
     3901 ggctcgggtg ccggattgcc ttcatttatg aaaattcccg gagaaaacct caacggagtt
     3961 tactcggcaa atgagtttct cacaagaata aacctcatga aggcttatga attccccaac
     4021 tgcgatactc ccgtgaaagt aggaaagaat gtcgccgttg tgggcggagg aaatgtcgca
     4081 atggacgccg caagaagcgc aaaaagactt ggcgcggaaa acgtttatat agtatacagg
     4141 cgttcggaag cggaaatgcc cgcaagactt gaagaaattc atcacgcaaa ggaagaagga
     4201 attttgttca aattccttac aaaccccaca agaattcttg gcaccgacga cggctgggtc
     4261 aaaggcatgg agtgcataga gatggagctg ggcgaacctg atgaatccgg aagaagaaga
     4321 cccgtgccaa agccgggatc cgaacatgta attgatgttg aaacggttat tatcgccatc
     4381 ggccaaactc caaatccgtt aattgcctca acaaccccgg ggctggccac tcaaaaatgg
     4441 ggcggaatta ttgtcgatga aaacaccggc gccaccaaca tagaaggtgt atatgccggc
     4501 ggagatgcgg taaccggtgc cgcaaccgtc attcttgcaa tgggagcagg caaaaaagcc
     4561 gcaaaggcaa ttgacgaata tcttaaaaac aaaaaatagt ttttaactgt aaaacaccat
     4621 ttaattgcgt tgtatatggt caaatgcaat aaatggtgtt tttgtctcta aaatttgatt
     4681 aactttaaac gcaaaaagca acgcttttaa aagcgttgct tttagaagta gccttaaggg
     4741 gagtcgaacc cctgtcttcg ccgtgagagg gcgatgtcct aggccactag acgataaggc
     4801 caaatattgg ctgccgaact aggattcgaa cctagactag atgatccaga gtcaccggtg
     4861 ctaccgttac accattcggc atcaactgac agaacttatt atagcatggg tttttatttt
     4921 tgacaagggg gtgttaaaaa atttttataa aaaactgact tgttttgagt gtttacatat
     4981 taagttaaat gctatataat atcctttgaa gttatatact gtatacataa aaggggggat
     5041 atgacaatgg cacatattat tacagacgaa tgcataagct gtggtgcttg cgctgcagaa
     5101 tgccctgtag atgctattca cgaaggtaca ggaaaatacg aagtggatgc tgatacatgt
     5161 atagcgtaaa acccagcgaa ccatttgagg tgataggtaa gattataccg aggtatgaaa
     5221 acgagaattg gacctttaca gaattactct atgaagcgcc atatttaaaa agctaccaag
     5281 acgaagagga tgaagaggat gaggaggcag attgccttga atatattgac aatactgata
     5341 agataatata tcttttatat agaagatatc gccgtatgta aggatttcag ggggcaaggc
     5401 ataggcagcg cgcttatcaa tatatctata gaatgggcaa agcataaaaa cttgcatgga
     5461 ctaatgcttg aaacccagga caataacctt atagcttgta aattctatca taattgtggt
     5521 ttcaaaatcg gctccgtcga tactatgtta tacgccaact ttcaaaacaa ctttgaaaaa
     5581 gctgttttct ggtatttaag gttttagaat gcaaggaaca gtgaattgga gttcgtcttg
     5641 ttataattag cttcttgggg tatctttaaa tactgtagaa aagaggaagg aaataataaa
     5701 tgaaaggacc aataataatg actagagaag aaagaatgaa gattgttcat gaaattaagg
     5761 aacgaatatt ggataaatat ggggatgatg ttaaggcaat tggtgtttat ggctctcttg
     5821 gtcgtcagac tgatgggccc tattcggata ttgagatgat gtgtgttctg tcaacagagg
     5881 gagtagagtt cagctatgaa tggacaaccg gtgagtggaa ggcggaagtg aatttttata
     5941 gcgaagagat tctactagat tatgcatctc gggtggaacc ggattggccg cttacacatg
     6001 gtcgattttt ctctattttg ccgatttatg atccaggtgg atactttgag aaagtgtacc
     6061 aaactgctaa atcggtagaa gcccaaaagt tccacgatgc gatctgtgcc cttatcgtag
     6121 aagagctgtt tgaatatgca ggcaaatggc gtaatattcg tgtgcaagga ccgacaacat
     6181 ttctaccatc cttgactgta caggtggcaa tggcaggtgc catgttgatt ggtctgcatc
     6241 atcgcatctg ttatacgacg agcgcttcgg tcttaactga agcagttaag caaccagatc
     6301 ttcctccagg ttatgtccaa ctgtgccagc tcgtaatgtc tggtcaactt tccgaccctg
     6361 agaaacttct ggaatcgcta gagaatttct ggaatggggt tcaggagtgg gcggaacgac
     6421 acggatatat agtggatgtg tcaaaacgca taccattttg atgacattaa gaaaggtggt
     6481 ttttatgata tacggttctt tggatcatgg ttttatagag gctatagttg gtcctatgtt
     6541 tagcggtaaa agtgaagaac tcataagaag gattaaaaga gcccaaattg ccaaacaaaa
     6601 agtacaagtc tttaaaccgg ctattgacga tagatattct attgacaagg tggtatctca
     6661 caatggcact aatataaatg ctattagtgt agtaaaggct tctgaaataa ttgagctttt
     6721 agaagaagac acagaggtta tcgcaattga tgaaattcag ttttttgacc attcgattgt
     6781 ggatgttgta agagaaattg cagatctagg caaaagagtc atttgtgctg gcttagatat
     6841 ggactttagg ggggaacctt ttggccctac tcctgatata atggcaattg cggagtctgt
     6901 agataaactg acggctatat gtgtgaaatg tggcaatcct gctactcgca ctcaaaggct
     6961 tatcaatggg aaacctgcaa aatacgatga cccaatcatt ttagtagggg cccatgaaac
     7021 ctatgaagca aggtgtcgaa aatgccatga agttcctcgt acatagtagt acctagattt
     7081 agatgcctaa aaagctttaa ctacaagctt tttagacatc taatcttttc tgaagtacat
     7141 ccgcaactgt ccatactaag aggcaaaaaa ggagaacctg tagctatagg cagactggag
     7201 agatttacag cagattggca tatgaaaaat aatgaagata aaatagaaaa accagagacg
     7261 aatggcagaa aagttgctgt tataggatct gggcctgcag gactatcgtg tgcaggtgat
     7321 cttgcaaaaa tgggatatga cacaaccatc tttgaagcat ttcatacacc aggcggtgtt
     7381 ttaatgtacg gtatcccaga gtttagactt ccaaaagaaa tagtgcaaaa agaaattgat
     7441 tcgttgaaaa aattgggagt aaaaatagaa accaacatgg tcataggcaa gatacttaca
     7501 atcgatgatt tattcgacat ggggtatgaa gcagtgttca tagggacagg cgcaggattg
     7561 cctaaattta tgaatatacc tggagaaaac ttaaatggag tttactcagc aaatgaattt
     7621 ttgacgagga ttaatttgat gaaagcttat gatttcccta acagtcccac tcctgtaaaa
     7681 gtaggcaaga aagttgctgt tgttggcggt g
//
