LOCUS       40924_20581        6377 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFT-A DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6377)
  AUTHORS   Okamoto S, Niki H.
  TITLE     cloning vector pFT-A sequence
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6377)
  AUTHORS   Okamoto S, Niki H.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-MAR-2016) Contact:Sho Okamoto National Institute of 
            Genetics, Microbial Genetics Laboratory, Genetics Strains Research 
            Center; 1111 Yata, Mishima, Shizuoka 411-8540, Japan URL 
            :https://www.nig.ac.jp/labs/MicroGen/
REFERENCE   3  (bases 1 to 6377)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6377)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (31-MAR-2016) Contact:Sho Okamoto National Institute of Genetics, 
            Microbial Genetics Laboratory, Genetics Strains Research Center; 
            1111 Yata, Mishima, Shizuoka 411-8540, Japan URL 
            :https://www.nig.ac.jp/labs/MicroGen/"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(8..1276)
                     /codon_start=1
                     /label=FLP
                     /note="site-specific recombinase"
                     /translation="MPQFGILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI
                     THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
                     IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
                     SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
                     KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
                     SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
                     YPAWNGIISQEVLDYLSSYINRRI"
     CDS             complement(1277..1564)
                     /codon_start=1
                     /label=SUMO
                     /note="cleavable ubiquitin-like protein tag"
                     /translation="DSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRR
                     LMEAFAKRQGKEMDSLTFLYDGIEIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             complement(1577..1594)
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     RBS             complement(1605..1627)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        1688..1716
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             1764..2951
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     CDS             complement(3544..4491)
                     /codon_start=1
                     /label=Rep101(Ts)
                     /note="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      complement(4539..4761)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(5386..6243)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6244..6348)
                     /label=AmpR promoter
ORIGIN
        1 tctattatat gcgtctattt atgtaggatg aaaggtagtc tagtacctcc tgtgatatta
       61 tcccattcca tgcggggtat cgtatgcttc cttcagcact accctttagc tgttctatat
      121 gctgccactc ctcaattgga ttagtctcat ccttcaatgc tatcatttcc tttgatattg
      181 gatcatatgc atagtaccga gaaactagtg cgaagtagtg atcaggtatt gctgttatct
      241 gatgagtata cgttgtcctg gccacggcag aagcacgctt atcgctccaa tttcccacaa
      301 cattagtcaa ctccgttagg cccttcattg aaagaaatga ggtcatcaaa tgtcttccaa
      361 tgtgagattt tgggccattt tttatagcaa agattgaata aggcgcattt ttcttcaaag
      421 ctttattgta cgatctgact aagttatctt ttaataattg gtattcctgt ttattgcttg
      481 aagaattgcc ggtcctattt actcgtttta ggactggttc agaattcctc aaaaattcat
      541 ccaaatatac aagtggatcg atcctacccc ttgcgctaaa gaagtatatg tgcctactaa
      601 cgcttgtctt tgtctctgtc actaaacact ggattattac tcccagatac ttattttgga
      661 ctaatttaaa tgatttcgga tcaacgttct taatatcgct gaatcttcca caattgatga
      721 aagtagctag gaagaggaat tggtataaag tttttgtttt tgtaaatctc gaagtatact
      781 caaacgaatt tagtattttc tcagtgatct cccagatgct ttcaccctca cttagaagtg
      841 ctttaagcat ttttttactg tggctatttc ccttatctgc ttcttccgat gattcgaact
      901 gtaattgcaa actacttaca atatcagtga tatcagattg atgtttttgt ccatagtaag
      961 gaataattgt aaattcccaa gcaggaatca atttctttaa tgaggcttcc agaattgttg
     1021 ctttttgcgt cttgtattta aactggagtg atttattgac aatatcgaaa ctcagcgaat
     1081 tgcttatgat agtattatag ctcatgaatg tggctctctt gattgctgtt ccgttatgtg
     1141 taatcatcca acataaatag gttagttcag cagcacataa tgctattttc tcacctgaag
     1201 gtctttcaaa cctttccaca aactgacgaa caagcacctt aggtggtgtt ttacataata
     1261 taccaaattg tggcatacct ccaatctgtt cgcggtgagc ctcaataata tcgttatcct
     1321 ccatgtccaa atcttcaggg gtctgatcag cttgaatttc aataccgtcg tacaagaacg
     1381 ttaaggagtc catttcctta ccctgtcttt tagcgaacgc ttccatcagc cttcttaaag
     1441 gagtggtctt tttgatcttg aagaagatct ctgaagatcc atcggacacc tttaaattga
     1501 tgtgagtctc aggcttgact tctggcttga cctctggctt agcttcttga ttgacttctg
     1561 agtcctgcag ggacccgtga tggtggtgat gatgcatatg tatatctcct tcttatagtt
     1621 aaacaaaatt atttctagtg tagcacctga agtcagcccc atacgatata agttgtaatt
     1681 ctcatgtttg acagcttatc atcgataagc tttaatgcgg tagtttatca cagttaaatt
     1741 gctaacgcag tcaggcaccg tgtatgaaat ctaacaatgc gctcatcgtc atcctcggca
     1801 ccgtcaccct ggatgctgta ggcataggct tggttatgcc ggtactgccg ggcctcttgc
     1861 gggatatcgt ccattccgac agcatcgcca gtcactatgg cgtgctgcta gcgctatatg
     1921 cgttgatgca atttctatgc gcacccgttc tcggagcact gtccgaccgc tttggccgcc
     1981 gcccagtcct gctcgcttcg ctacttggag ccactatcga ctacgcgatc atggcgacca
     2041 cacccgtcct gtggatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag
     2101 gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact
     2161 tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac
     2221 tgttgggcgc catctccttg catgcaccat tccttgcggc ggcggtgctc aacggcctca
     2281 acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt cgaccgatgc
     2341 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     2401 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     2461 gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg
     2521 cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac
     2581 gtttcggcga gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct
     2641 tgctggcgtt cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg
     2701 gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc
     2761 agggacagct tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc attggaccgc
     2821 tgatcgtcac ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg
     2881 taggcgccgc cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg
     2941 ccacctcgac ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat
     3001 tggagccaat caattcttgc ggagaactgt gaatgcgcaa cccaaccctt ggcagaacat
     3061 atccatcgcg tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc
     3121 ctggccacgg gtgcgcatga tcgtgctcct gtcgctagat aatcgtccat tccgacagca
     3181 tcgccagtca ctatggcgtg ctgctagcgc tatatgcgtt gatgcaattt ctatgcgcac
     3241 ccgttctcgg agcactgtcc gaccgctttg gccgccgccc agtcctgctc gcttcgctac
     3301 ttggagccac tatcgactac gcgatcatgg cgaccacacc cgtcctgtgg atccgctagg
     3361 gataacaggg taatatagat cctctagagt cgacctgcag gcatgcaagc ttggcactgg
     3421 ctgatcagct agcccatggg tatggacagt tttccctttg atatgtaacg gtgaacagtt
     3481 gttctacttt tgtttgttag tcttgatgct tcactgatag atacaagagc cataagaacc
     3541 tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag
     3601 ccatgagaac gaaccattga gatcatgctt actttgcatg tcactcaaaa attttgcctc
     3661 aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt
     3721 acgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg
     3781 gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa
     3841 cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa
     3901 atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt
     3961 ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt
     4021 tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc
     4081 aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg
     4141 caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt
     4201 ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat
     4261 cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat
     4321 ctgaacgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat
     4381 cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc
     4441 atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa
     4501 ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat gataattact
     4561 agtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact
     4621 tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta
     4681 tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga
     4741 tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc
     4801 gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac
     4861 cctcgcaagc tcggttgcgg ccgcaatcgg gcaaatcgct gaatattcct tttgtctccg
     4921 accatcaggc acctgagtcg ctgtcttttt cgtgacattc agttcgctgc gctcacggct
     4981 ctggcagtga atgggggtaa atggcactac aggcgccttt tatggattca tgcaaggaaa
     5041 ctacccataa tacaagaaaa gcccgtcacg ggcttctcag ggcgttttat ggcgggtctg
     5101 ctatgtggtg ctatctgact ttttgctgtt cagcagttcc tgccctctga ttttccagtc
     5161 tgaccacttc ggattatccc gtgacaggtc attcagactg gctaatgcac ccagtaaggc
     5221 agcggtatca tcaacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5281 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5341 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5401 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5461 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5521 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5581 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5641 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5701 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5761 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5821 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5881 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5941 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6001 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6061 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6121 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6181 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6241 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6301 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6361 aaaagtgcca cttgcat
//
