LOCUS       Exported                7454 bp ds-DNA     circular SYN 18-OCT-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7454)
  AUTHORS   gep
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Oct 21, 2019 from SnapGene Viewer 5.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7454
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(1..879)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     misc_feature    complement(891..1020)
                     /label=promoter for AraC
     promoter        1025..1190
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction"
     misc_feature    1225..1938
                     /label=I-SceI endonuclease
     misc_feature    1915..1922
                     /label=UAS repeats
     CDS             1979..2395
                     /codon_start=1
                     /label=gam protein
                     /translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
                     IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
                     AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
     CDS             2401..3186
                     /codon_start=1
                     /label=bet protein
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     CDS             3183..3863
                     /codon_start=1
                     /label=exonuclease
                     /translation="MTPDIILQRTGIDVRAVEQGEDAWHKLRLGVITASEVHNVIAKPR
                     SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     CDS             complement(4361..5311)
                     /codon_start=1
                     /gene="rep101 encoding an A56V mutation"
                     /product="temperature-sensitive version of a protein needed
                     for replication with the pSC101 origin"
                     /label=Rep101(Ts)
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      5359..5581
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             6375..7190
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac ggaactcgct
       61 cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc
      121 aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg
      181 gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct ggcggaaaag
      241 atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga tatcaaaatt
      301 gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat tatccatcgg
      361 tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct caagcagatt
      421 tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga tttgcccaaa
      481 caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg tattggcaaa
      541 tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt aaacccactg
      601 gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc ctggcgggaa
      661 cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg
      721 accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt cgataaaaaa
      781 atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg cattaaacga
      841 gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac tcccgccatt
      901 cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg tcttttactg
      961 gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt aacaaagcgg
     1021 gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca
     1081 cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat ccataagatt
     1141 agcggatcct acctgacgct ttttatcgca actctctact gtttctccat acccgttttt
     1201 ttgggctagc aggagggtac ctatatgcat atgaaaatca tcaaaaaaaa ccaggtaatg
     1261 aacctcggtc cgaactctaa actgctgaaa gaatacaaat cccagctgat cgaactgaac
     1321 atcgaacagt tcgaagcagg tatcggtctg atcctgggtg atgcttacat ccgttctcgt
     1381 gatgaaggta aaacctactg tatgcagttc gagtggaaaa acaaagcata catggaccac
     1441 gtatgtctgc tgtacgatca gtgggtactg tccccgccgc acaaaaaaga acgtgttaac
     1501 cacctgggta acctggtaat cacctggggc gcccagactt tcaaacacca agcattcaac
     1561 aaactggcta acctgttcat cgttaacaac aaaaaaacca tcccgaacaa cctggttgaa
     1621 aactacctga ccccgatgtc tctggcatac tggttcatgg atgatggtgg taaatgggat
     1681 tacaacaaaa actctaccaa caaatcgatc gtactgaaca cccagtcttt cactttcgaa
     1741 gaagtagaat acctggttaa gggtctgcgt aacaaattcc aactgaactg ttacgtaaaa
     1801 atcaacaaaa acaaaccgat catctacatc gattctatgt cttacctgat cttctacaac
     1861 ctgatcaaac cgtacctgat cccgcagatg atgtacaaac tgccgaacac tatctcctcc
     1921 gaaactttcc tgaaataagc tagaagatgt ttcgtgaagc cgtcgacgct tataaaaaat
     1981 ggatattaat actgaaactg agatcaagca aaagcattca ctaaccccct ttcctgtttt
     2041 cctaatcagc ccggcatttc gcgggcgata ttttcacagc tatttcagga gttcagccat
     2101 gaacgcttat tacattcagg atcgtcttga ggctcagagc tgggcgcgtc actaccagca
     2161 gctcgcccgt gaagagaaag aggcagaact ggcagacgac atggaaaaag gcctgcccca
     2221 gcacctgttt gaatcgctat gcatcgatca tttgcaacgc cacggggcca gcaaaaaatc
     2281 cattacccgt gcgtttgatg acgatgttga gtttcaggag cgcatggcag aacacatccg
     2341 gtacatggtt gaaaccattg ctcaccacca ggttgatatt gattcagagg tataaaacga
     2401 atgagtactg cactcgcaac gctggctggg aagctggctg aacgtgtcgg catggattct
     2461 gtcgacccac aggaactgat caccactctt cgccagacgg catttaaagg tgatgccagc
     2521 gatgcgcagt tcatcgcatt actgatcgtt gccaaccagt acggccttaa tccgtggacg
     2581 aaagaaattt acgcctttcc tgataagcag aatggcatcg ttccggtggt gggcgttgat
     2641 ggctggtccc gcatcatcaa tgaaaaccag cagtttgatg gcatggactt tgagcaggac
     2701 aatgaatcct gtacatgccg gatttaccgc aaggaccgta atcatccgat ctgcgttacc
     2761 gaatggatgg atgaatgccg ccgcgaacca ttcaaaactc gcgaaggcag agaaatcacg
     2821 gggccgtggc agtcgcatcc caaacggatg ttacgtcata aagccatgat tcagtgtgcc
     2881 cgtctggcct tcggatttgc tggtatctat gacaaggatg aagccgagcg cattgtcgaa
     2941 aatactgcat acactgcaga acgtcagccg gaacgcgaca tcactccggt taacgatgaa
     3001 accatgcagg agattaacac tctgctgatc gccctggata aaacatggga tgacgactta
     3061 ttgccgctct gttcccagat atttcgccgc gacattcgtg catcgtcaga actgacacag
     3121 gccgaagcag taaaagctct tggattcctg aaacagaaag ccgcagagca gaaggtggca
     3181 gcatgacacc ggacattatc ctgcagcgta ccgggatcga tgtgagagct gtcgaacagg
     3241 gggaagatgc gtggcacaaa ttacggctcg gcgtcatcac cgcttcagaa gttcacaacg
     3301 tgatagcaaa accccgctcc ggaaagaagt ggcctgacat gaaaatgtcc tacttccaca
     3361 ccctgcttgc tgaggtttgc accggtgtgg ctccggaagt taacgctaaa gcactggcct
     3421 ggggaaaaca gtacgagaac gacgccagaa ccctgtttga attcacttcc ggcgtgaatg
     3481 ttactgaatc cccgatcatc tatcgcgacg aaagtatgcg taccgcctgc tctcccgatg
     3541 gtttatgcag tgacggcaac ggccttgaac tgaaatgccc gtttacctcc cgggatttca
     3601 tgaagttccg gctcggtggt ttcgaggcca taaagtcagc ttacatggcc caggtgcagt
     3661 acagcatgtg ggtgacgcga aaaaatgcct ggtactttgc caactatgac ccgcgtatga
     3721 agcgtgaagg cctgcattat gtcgtgattg agcgggatga aaagtacatg gcgagttttg
     3781 acgagatcgt gccggagttc atcgaaaaaa tggacgaggc actggctgaa attggttttg
     3841 tatttgggga gcaatggcga tgacgcatcc tcacgataat aagcttcctg ctgaacatca
     3901 aaggcaagaa aacatctgtt gtcaaagaca gcatccttga acaaggacaa ttaacagtta
     3961 acaaataaaa acgcaaaaga aaatgccgat attgactacc ggaagcagtg tgaccgtgtg
     4021 cttctcaaat gcctgattca ggctgtctat gtgtgactgt tgagctgtaa caagttgtct
     4081 caggtgttca atttcatgtt ctagttgctt tgttttactg gtttcacctg ttctattagg
     4141 tgttacatgc tgttcatctg ttacattgtc gatctgttca tggtgaacag ctttaaatgc
     4201 accaaaaact cgtaaaagct ctgatgtatc tatctttttt acaccgtttt catctgtgca
     4261 tatggacagt tttccctttg atatgtaacg gtgaacagtt gttctacttt tgtttgttag
     4321 tcttgatgct tcactgatag atacaagagc cataagaacc tcagatcctt ccgtatttag
     4381 ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac gaaccattga
     4441 gatcatactt actttgcatg tcactcaaaa attttgcctc aaaactggtg agctgaattt
     4501 ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt atgtaggtag gaatctgatg
     4561 taatggttgt tggtattttg tcaccattca tttttatctg gttgttctca agttcggtta
     4621 cgagatccat ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc
     4681 gcttatcaac caccaatttc atattgctgt aagtgtttaa atctttactt attggtttca
     4741 aaacccattg gttaagcctt ttaaactcat ggtagttatt ttcaagcatt aacatgaact
     4801 taaattcatc aaggctaatc tctatatttg ccttgtgagt tttcttttgt gttagttctt
     4861 ttaataacca ctcataaatc ctcatagagt atttgttttc aaaagactta acatgttcca
     4921 gattatattt tatgaatttt tttaactgga aaagataagg caatatctct tcactaaaaa
     4981 ctaattctaa tttttcgctt gagaacttgg catagtttgt ccactggaaa atctcaaagc
     5041 ctttaaccaa aggattcctg atttccacag ttctcgtcat cagctctctg gttgctttag
     5101 ctaatacacc ataagcattt tccctactga tgttcatcat ctgaacgtat tggttataag
     5161 tgaacgatac cgtccgttct ttccttgtag ggttttcaat cgtggggttg agtagtgcca
     5221 cacagcataa aattagcttg gtttcatgct ccgttaagtc atagcgacta atcgctagtt
     5281 catttgcttt gaaaacaact aattcagaca tacatctcaa ttggtctagg tgattttaat
     5341 cactatacca attgagatgg gctagtcaat gataattact agtccttttc ctttgagttg
     5401 tgggtatctg taaattctgc tagacctttg ctggaaaact tgtaaattct gctagaccct
     5461 ctgtaaattc cgctagacct ttgtgtgttt tttttgttta tattcaagtg gttataattt
     5521 atagaataaa gaaagaataa aaaaagataa aaagaataga tcccagccct gtgtataact
     5581 cactacttta gtcagttccg cagtattaca aaaggatgtc gcaaacgctg tttgctcctc
     5641 tacaaaacag accttaaaac cctaaaggct taagtagcac cctcgcaagc tcgggcaaat
     5701 cgctgaatat tccttttgtc tccgaccatc aggcacctga gtcgctgtct ttttcgtgac
     5761 attcagttcg ctgcgctcac ggctctggca gtgaatgggg gtaaatggca ctacaggcgc
     5821 cttttatgga ttcatgcaag gaaactaccc ataatacaag aaaagcccgt cacgggcttc
     5881 tcagggcgtt ttatggcggg tctgctatgt ggtgctatct gactttttgc tgttcagcag
     5941 ttcctgccct ctgattttcc agtctgacca cttcggatta tcccgtgaca ggtcattcag
     6001 actggctaat gcacccagta aggcagcggt atcatcaaca ggcttacccg tcttactgtc
     6061 ggggatcgac gctctccctt atgcgactcc tgcacctttc gtcttcgaat aaatacctgt
     6121 gacggaagat cacttcgcag ataaaataaa tcctggagtt ggtagctctt gatccggcaa
     6181 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6241 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6301 aaactcacgt taagggattt tggtcatgaa caataaaact gtctgcttac ataaacagta
     6361 atacaagggg tgttatgagc catattcaac gggaaacgtc ttgctctagg ccgcgattaa
     6421 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
     6481 caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac
     6541 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
     6601 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt
     6661 tactcaccac tgcgatcccc gggaaaacag cattccaggt attagaagaa tatcctgatt
     6721 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg
     6781 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
     6841 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
     6901 aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc
     6961 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     7021 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     7081 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     7141 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa gaattaattc
     7201 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7261 tttccccgaa aagtgccacc tgtcgtcggt tcagggcagg gtcgttaaat agccgcttat
     7321 gtctattgct ggtctcggta cccggggatc ctctagagtc gacctgcata atgtgcctgt
     7381 caaatggacg aagcagggat tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc
     7441 tgattcgtta ccaa
//
