LOCUS       V015919                 8625 bp    DNA     circular SYN 10-MAR-2019
DEFINITION  Exported.
ACCESSION   V015919
VERSION     V015919
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8625)
  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 8625)
  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..8625
                     /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..2415
                     /label="upstream homology recombination flank"
                     /note="upstream homology recombination flank"
     CDS             2416..2994
                     /codon_start=1
                     /transl_table=11
                     /product="pyruvate ferredoxin oxidoreductase gamma subunit"
                     /EC_number="1.2.7.1"
                     /label="pyruvate ferredoxin oxidoreductase gamma subunit"
                     /note="Clo1313_0020"
                     /protein_id="QBI89806.1"
                     /translation="MGKVVEIRWHGRGGQGAKTASLLLADAAFNTGKYIQGFPEYGPER
                     MGAPITAYNRISDEKLTIHSNIYEPDYVVVVDDTLLTSVDVTAGLKEDGAIIVNTPKTP
                     DEIRPLLKGYKGKVCTIDARKISIETLGKYFPNTPMLGAVVKVSKIMDEEEFLKDMVES
                     FKHKFANKPEVVEGNIKALERSMQEVKGL"
     CDS             2991..3296
                     /codon_start=1
                     /transl_table=11
                     /product="pyruvate ferredoxin oxidoreductase delta subunit"
                     /EC_number="1.2.7.1"
                     /label="pyruvate ferredoxin oxidoreductase delta subunit"
                     /note="Clo1313_0021"
                     /protein_id="QBI89807.1"
                     /translation="MSKELRDVKPDVTWKEITSGGVIDSPGNAHLFKTGDWRSMKPVWN
                     EEKCKQCLLCNPVCPDSSIMVSEEGKMTGIDYDHCKGCGICSKVCPFKAIDFVEEV"
     CDS             3313..4497
                     /codon_start=1
                     /transl_table=11
                     /product="pyruvate ferredoxin oxidoreductase alpha subunit"
                     /EC_number="1.2.7.1"
                     /label="pyruvate ferredoxin oxidoreductase alpha subunit"
                     /note="Clo1313_0022"
                     /protein_id="QBI89808.1"
                     /translation="MGIRERLSGNEATAIAMRQINPDVVAAFPITPSTEIPQYFSSYVA
                     DGLVDTEFVAVESEHSAMSACIGAQAAGARAMTATSANGLAYMWEALYIAASMRLPIVL
                     AAVNRALSGPINIHNDHSDTMGARDSGWIQLYSENNQEAYDNMLMAHRIGEHPDVMLPV
                     MVCQDGFITSHAIENIELVEDEKVKAFVGEYKPTHYLLDRENPISVGPLDLQMHYFEHK
                     RQQAQAMENAKKVILEVAEEFYKLTGRKYGFFEEYKTDDADVAIVVMNSTAGTVKYVID
                     EYRVKGKKVGLIKPRVFRPFPVDELAQALSKFKAVAVMDKADSFNAAGGPLFTEVTSAL
                     FTKGVFGPKVINYKFGLGGRDVKVDDIEVVCEKLLEIASTGKVDSVYNYLGVRE"
     CDS             4510..5445
                     /codon_start=1
                     /transl_table=11
                     /product="pyruvate ferredoxin oxidoreductase beta subunit"
                     /EC_number="1.2.7.1"
                     /label="pyruvate ferredoxin oxidoreductase beta subunit"
                     /note="Clo1313_0023"
                     /protein_id="QBI89809.1"
                     /translation="MAYNLKEVAKKPERLTGGHRMCAGCGAPIVVRQVLKALKPEDHAV
                     ISAATGCLEVSTFIYPYTAWKDSFIHSAFENTGATISGAEAAYKVLKKKGKIEGETKFI
                     AFGGDGGTYDIGLQALSGAMERGHDMVYVCYDNGAYMNTGIQRSSATPKYADTTTSPVG
                     KKIPGKMQPRKDLTEVLVNHRIPYVAQTAPFGNMKDLYEKAEKAIYTPGPAFLNVLAPC
                     PRGWRYNTPDLMELSKLAVETCFWPLYEVIDGKYIINYKPKEKVPVKEFLKLQGRFKHL
                     FKAGNEYMLEEIQKEVDLRWERLLKLAGEA"
     misc_feature    5446..5988
                     /label="downstream homology recombination flank"
                     /note="downstream homology recombination flank"
     regulatory      5993..6523
                     /label="kanamycin promoter"
                     /note="kanamycin promoter"
                     /regulatory_class="promoter"
     gene            6524..7285
                     /gene="htk"
                     /label="htk"
     CDS             6524..7285
                     /codon_start=1
                     /transl_table=11
                     /gene="htk"
                     /product="highly thermostable kanamycin
                     nucleotidyltransferase"
                     /EC_number="2.7.7.46"
                     /label="htk"
                     /protein_id="QBI89810.1"
                     /translation="MKGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGRQTD
                     GPYSDIEMMCVLSTEGVEFSYEWTTGEWKAEVNFYSEEILLDYASRVEPDWPLTHGRFF
                     SILPIYDPGGYFEKVYQTAKSVEAQKFHDAICALIVEELFEYAGKWRNIRVQGPTTFLP
                     SLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQPDLPPGYVQLCQLVMSGQLSDPEK
                     LLESLENFWNGVQEWAERHGYIVDVSKRIPF"
     CDS             7309..7887
                     /gene="tdk"
                     /label="Thymidine kinase"
                     /note="Thymidine kinase from Thermoanaerobacter
                     pseudethanolicus (strain ATCC 33223 / 39E). Accession#:
                     B0K7G9"
     misc_feature    7982..8625
                     /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 ggttctatct cattcctttc
     1861 acacgatatg aaaatttttg aaataataga cgtgatatta tgcagaattt taatatttga
     1921 aatattttat aaaagccact atctatctat aaattagata aatactggct aagcggtaaa
     1981 tttgcaaagc aaccctttca tagaaacccc ctatttactt tccttcaatg tagaaaaaat
     2041 tcatgtaatt tattttatca ttttcaagca ttaaagtcaa tccctgctta acaaaataca
     2101 caattagccg aaatattaaa ttatagatgc tgtggcaact taacaatatt atatcacata
     2161 atatgaccag ctcataattt ttatgaataa atttttgtca aaaaaatatt tttatatttt
     2221 aattatctgt gatataatta tcgtggaact gttatataaa gtctaaaggt atatatataa
     2281 gcattttcag aaaactgtta tataatcgct taaccgctgt tgtataacag tacatatttg
     2341 tttatgctgt attataatag tttgcttata tatccttaga tttaaatata aatttaaaaa
     2401 gagaggggaa atacaatggg caaggttgtt gaaatcagat ggcacgggcg cggcggacaa
     2461 ggtgccaaga ctgcgtcgct tctgctggct gatgctgcat tcaataccgg aaaatacatc
     2521 cagggttttc cggaatacgg tcctgaaaga atgggcgcgc caattacagc ttacaacagg
     2581 ataagtgatg agaaacttac cattcacagc aatatttatg aaccggatta tgttgtggtt
     2641 gtagatgaca ctcttttgac atccgttgat gttacagcag gtttgaaaga ggatggagct
     2701 attatcgtca acactccaaa aactcctgac gaaataagac ctcttttaaa aggctacaag
     2761 ggaaaggttt gcacaattga cgcaagaaaa atttccattg aaactttggg aaaatacttc
     2821 ccgaacaccc caatgcttgg tgctgttgta aaggttagca agataatgga cgaagaagaa
     2881 ttcctcaagg atatggttga gtcattcaag cacaagtttg cgaacaagcc tgaagttgtt
     2941 gaaggcaata tcaaggccct ggagaggtca atgcaggagg tgaaaggatt atgagtaaag
     3001 aattgagaga tgtaaaacct gatgttacct ggaaagagat aacatccggt ggagtgatag
     3061 attcccctgg aaacgcacat ctttttaaga ccggtgactg gcgttctatg aagccggtgt
     3121 ggaatgagga aaaatgcaag cagtgtttgc tttgcaaccc ggtatgtcct gattcatcca
     3181 taatggtaag tgaagaggga aaaatgacag gtatcgatta tgatcactgc aaaggatgtg
     3241 gcatttgttc aaaggtttgt cctttcaaag ccatagactt tgtagaggaa gtataagaga
     3301 gggggacaca aaatgggaat aagagaaagg cttagcggta atgaggcaac ggcgattgcc
     3361 atgagacaaa taaatcctga tgtggttgct gcttttccga taacaccgtc aacggaaatt
     3421 cctcaatatt tctcgtcata tgtcgctgac ggacttgtag atacggaatt tgttgctgtg
     3481 gaatcagagc acagtgcaat gtctgcatgt ataggtgctc aggctgcagg tgcaagagca
     3541 atgactgcca catccgcaaa cggtttggca tatatgtggg aggctttgta tatagcggca
     3601 agtatgagac ttccgattgt attggcggct gtaaacagag cactttcagg tcctatcaat
     3661 atccacaacg accacagcga tacaatggga gctagggatt cgggatggat ccagttatac
     3721 agtgaaaaca accaggaggc ttatgacaac atgcttatgg ctcacaggat aggtgagcat
     3781 cctgatgtaa tgcttcctgt catggtctgc caggacggat ttattacttc tcacgcaata
     3841 gaaaatattg aactggtgga agatgagaaa gttaaggctt ttgtaggaga atacaaaccg
     3901 actcattatc ttctcgacag ggaaaatccg atttctgtgg gtcctttgga tttgcagatg
     3961 cattatttcg agcacaagag acagcaggca caggcaatgg aaaacgccaa aaaggtaatt
     4021 cttgaagtgg cggaagaatt ctacaagctt acgggaagaa aatacggatt ttttgaagaa
     4081 tacaaaaccg atgatgccga tgttgccatt gttgttatga actccactgc cggtactgta
     4141 aaatatgtta tcgacgagta cagggtaaaa ggcaaaaaag ttggtttgat aaaacctaga
     4201 gtattcagac ctttccctgt tgatgaactg gcacaggctt tgtcaaagtt taaggcagtg
     4261 gccgttatgg acaaggctga cagcttcaat gcagccggag gacctttgtt tacagaggta
     4321 acaagtgcac tcttcacaaa aggagtattt ggtcctaagg ttattaacta taagtttgga
     4381 ttgggtggaa gagacgttaa agttgatgat attgaagttg tttgtgagaa gcttctggaa
     4441 attgcaagta caggcaaggt agactcagta tacaattacc ttggtgttag agagtaggag
     4501 gtgtggtaaa tggcttataa tttgaaagaa gttgcaaaaa aaccggaaag acttacgggc
     4561 ggacacagaa tgtgtgcagg ctgcggagct ccgatagttg taagacaggt tcttaaggca
     4621 ttaaaaccgg aagatcatgc ggttatctca gctgcaacag gttgtttgga agtttcaact
     4681 ttcatttacc cttatacagc atggaaggat tctttcattc acagtgcgtt tgaaaatacc
     4741 ggtgctacaa tttccggtgc ggaagcggct tataaagtat tgaagaaaaa aggaaaaatt
     4801 gaaggggaga ccaagtttat tgcgttcggt ggtgacggcg gaacatacga cataggactt
     4861 caggcactct caggagcgat ggaaagagga cacgacatgg tttatgtgtg ctacgacaat
     4921 ggagcataca tgaacacagg tatccagagg tcttctgcca ctccgaaata cgctgatacc
     4981 acaacttcac ctgttggaaa gaagataccc ggtaaaatgc agccaagaaa agacctgaca
     5041 gaagtattgg taaatcatcg cataccttat gttgctcaaa ccgctccttt cgggaacatg
     5101 aaggacctct atgaaaaagc tgaaaaagct atttatacac ccggtcctgc gttcctgaac
     5161 gtgttggcac cgtgcccgag aggatggaga tacaacactc ctgatttgat ggaattgagc
     5221 aaattggcgg ttgaaacttg cttctggccg ctttatgaag taattgacgg caaatatata
     5281 ataaactaca agccgaagga aaaagttccc gtcaaggaat tcttgaaact tcagggaaga
     5341 tttaaacatc ttttcaaagc cggcaacgaa tatatgctgg aagaaattca gaaagaagtc
     5401 gacttaagat gggagagact cttgaagctg gccggagagg cttaacttca ccacaattaa
     5461 tgagcgaaat tgctttcgct cattaatttt ttatgtgtta gttgcctcct tctaaatttt
     5521 tctcttgttt atctatttaa tgttttatat aattaatgtt ataaactaca taaaaggagc
     5581 ggatgatatg agagaaaaaa taaagatggc aaatcctatc gtagagctgg atggcgatga
     5641 aatgaccaga atcatctggc agatgataaa agacacgctt atacatcctt ttgtagaact
     5701 gaatacagaa tactacgatc tgggtcttaa aaatagagat ttgacagacg accaagtgac
     5761 tgtagatgcc gcctatgcga taaaaaagca caaagtaggc gtcaaatgcg caactataac
     5821 acctaatgct caaagagtcg aagaatacaa cttaaaaaag atgtggaaaa gccccaatgg
     5881 caccataagg gctatattgg acggcaccgt ttttaggact ccaatactgg taaacggcat
     5941 caaacctctc gtaaaaagct ggacaaagcc tataacgatt gcgcgtcacg taaaacccag
     6001 cgaaccattt gaggtgatag gtaagattat accgaggtat gaaaacgaga attggacctt
     6061 tacagaatta ctctatgaag cgccatattt aaaaagctac caagacgaag aggatgaaga
     6121 ggatgaggag gcagattgcc ttgaatatat tgacaatact gataagataa tatatctttt
     6181 atatagaaga tatcgccgta tgtaaggatt tcagggggca aggcataggc agcgcgctta
     6241 tcaatatatc tatagaatgg gcaaagcata aaaacttgca tggactaatg cttgaaaccc
     6301 aggacaataa ccttatagct tgtaaattct atcataattg tggtttcaaa atcggctccg
     6361 tcgatactat gttatacgcc aactttcaaa acaactttga aaaagctgtt ttctggtatt
     6421 taaggtttta gaatgcaagg aacagtgaat tggagttcgt cttgttataa ttagcttctt
     6481 ggggtatctt taaatactgt agaaaagagg aaggaaataa taaatgaaag gaccaataat
     6541 aatgactaga gaagaaagaa tgaagattgt tcatgaaatt aaggaacgaa tattggataa
     6601 atatggggat gatgttaagg caattggtgt ttatggctct cttggtcgtc agactgatgg
     6661 gccctattcg gatattgaga tgatgtgtgt tctgtcaaca gagggagtag agttcagcta
     6721 tgaatggaca accggtgagt ggaaggcgga agtgaatttt tatagcgaag agattctact
     6781 agattatgca tctcgggtgg aaccggattg gccgcttaca catggtcgat ttttctctat
     6841 tttgccgatt tatgatccag gtggatactt tgagaaagtg taccaaactg ctaaatcggt
     6901 agaagcccaa aagttccacg atgcgatctg tgcccttatc gtagaagagc tgtttgaata
     6961 tgcaggcaaa tggcgtaata ttcgtgtgca aggaccgaca acatttctac catccttgac
     7021 tgtacaggtg gcaatggcag gtgccatgtt gattggtctg catcatcgca tctgttatac
     7081 gacgagcgct tcggtcttaa ctgaagcagt taagcaacca gatcttcctc caggttatgt
     7141 ccaactgtgc cagctcgtaa tgtctggtca actttccgac cctgagaaac ttctggaatc
     7201 gctagagaat ttctggaatg gggttcagga gtgggcggaa cgacacggat atatagtgga
     7261 tgtgtcaaaa cgcataccat tttgatgaca ttaagaaagg tggtttttat gatatacggt
     7321 tctttggatc atggttttat agaggctata gttggtccta tgtttagcgg taaaagtgaa
     7381 gaactcataa gaaggattaa aagagcccaa attgccaaac aaaaagtaca agtctttaaa
     7441 ccggctattg acgatagata ttctattgac aaggtggtat ctcacaatgg cactaatata
     7501 aatgctatta gtgtagtaaa ggcttctgaa ataattgagc ttttagaaga agacacagag
     7561 gttatcgcaa ttgatgaaat tcagtttttt gaccattcga ttgtggatgt tgtaagagaa
     7621 attgcagatc taggcaaaag agtcatttgt gctggcttag atatggactt taggggggaa
     7681 ccttttggcc ctactcctga tataatggca attgcggagt ctgtagataa actgacggct
     7741 atatgtgtga aatgtggcaa tcctgctact cgcactcaaa ggcttatcaa tgggaaacct
     7801 gcaaaatacg atgacccaat cattttagta ggggcccatg aaacctatga agcaaggtgt
     7861 cgaaaatgcc atgaagttcc tcgtacatag tagtacctag atttagatgc ctaaaaagct
     7921 ttaactacaa gctttttaga catctaatct tttctgaagt acatccgcaa ctgtccatac
     7981 tacatcgcta agaacaacat caatttctac atcataaatg cagttgacat agcaaaagaa
     8041 ttaggtcttg gcgcaagaat aaacatgata atgcaatcag cattctttaa gcttgcgaac
     8101 atcattccta ttgatgaagc agttaaacac ttaaaagatg ctattgtgaa gtcttacggt
     8161 cataaaggcg aaaagatagt caacatgaac tacgcagccg tcgatcgtgg aatagatgca
     8221 cttgttaagg tagatgttcc tgcatcgtgg gcaaatgctg aagacgaagc taaagtagaa
     8281 agaaatgttc ctgatttcat aaagaatata gcagatgtta tgaatagaca agaaggagac
     8341 aaattgcctg taagcgcctt tgtaggcatg gaagacggta cattcccaat gggcactgca
     8401 gcatacgaga agcgcggtat cgccgtagat gtgcctgaat ggcagataga taactgcata
     8461 cagtgcaatc agtgtgcgta tgtatgtcca catgcagcaa tcaggccatt cctcttaaat
     8521 gaagaagaag tcaaaaacgc tcctgaagga tttacttcca aaaaagctat cggtaagggc
     8581 ttagaaggat tgaacttcag gattcaagtg agcgtgcttg attgt
//
