LOCUS       Exported                8187 bp ds-DNA     circular SYN 15-MAR-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8187)
  AUTHORS   StruBiol
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Mar 24, 2021 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8187
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(183..771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(942..1802)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1803..1907)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2379..2395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2410..3124
                     /label=arg3 upstream sequence
     misc_feature    2410..2896
                     /label=targeting sequence L
     gene            3125..4108
                     /label=arg3 CDS
     misc_feature    4109..4666
                     /label=arg3 downstream sequence
     promoter        4697..5438
                     /label=adh1 promoter
                     /note="promoter for the Schizosaccharomyces pombe adh1 gene
                     encoding alcohol dehydrogenase"
     CDS             5496..7253
                     /codon_start=1
                     /label=OsTIR1+
                     /translation="MTYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFV
                     GNCYAVRAGRVAARFPNVRALTVKGKPHFADFNLVPPDWGGYAGPWIEAAARGCHGLEE
                     LRMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEV
                     EDRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTL
                     AKILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYP
                     LCAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELR
                     VFPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRL
                     CILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSI
                     AFAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKG
                     CQVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKILG
                     SGPKKKRKV"
     CDS             7230..7250
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      7272..7459
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     misc_feature    7495..7941
                     /label=targeting sequence R
     CDS             complement(7518..7941)
                     /codon_start=1
                     /label=cmb1
                     /translation="ALTADEIQKYKNKLQEQFDAEKKRFMETLRSFTPTEIDSENRRRS
                     KEAHSTGSRYYRLRHPDVPKKPSSAFILFYKELRNNPKLRQELGIPEAISTLVEETQNA
                     SKAWKELAEDKKKPFIDKSKALKEQYDKFMKEAGFR"
     primer_bind     complement(7968..7984)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7992..8008
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(8016..8046)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8061..8082
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
       61 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      121 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      181 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      241 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      301 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      361 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      421 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      481 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      541 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      601 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      661 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      721 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      781 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      841 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      901 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      961 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1021 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1081 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1141 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1201 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1261 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1321 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1381 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1441 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1501 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1561 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1621 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1681 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1741 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1801 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1861 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     1921 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     1981 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     2041 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     2101 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     2161 gagcagattg tactgagagt gcaccatgcg gtgtgaaata ccgcacagat gcgtaaggag
     2221 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     2281 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     2341 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     2401 cgcggccgca gcttgtaata tagcgaatct gtgaactcat tgttcatttc atggtagttg
     2461 ttgatttcac aagcgtgctg agttatcaat atagtaacac aattggtttt atttcagctc
     2521 gcttgaatat agtgtcaaga gataattgaa taaacaaaaa aaataactac acgtttttta
     2581 gatacatttt atttacaatg tattggttct ttaaagaatg ggtggtaaat aaaaaaaata
     2641 ttacaaggtc gaaaaaaatt catccctttt cattagttct ataattaaat gctaaaccca
     2701 tgaatattct tttactgcta atcaattaac gattttggta tttaaaagct attgctgata
     2761 agtattctga cgttctgtat caatattacc atgaataaca taagaaaaag aaaggaaata
     2821 gaatgtttat gagtagtttt taatttttga cttgttttat catatttcat caacgtacta
     2881 gcttgtttgc aactgaactc tagtgtacga taatgtaaaa attttccatc tcgtcgaaaa
     2941 tccgcatacg atatattacg gcgccaatct cgcagtctga gagagaacta gatatcggct
     3001 atatgcaatc tcaccctgaa tatctaagaa attaccagta acttgtgggt tccttgtagt
     3061 cgtctacaaa aaccgtattt ttgatatatt tagtcgagaa gaattattca gtttgcaatt
     3121 gaaaatgtct ttcaaaaaat ttcctcgtca tttactatcc attcgtgatc ttagtcgcgg
     3181 agagattgta aagttgatcg atcggtcttc tgaaattaaa caagcctata agcagaattt
     3241 tcagaatcgc cgttctgttc aaatgagtgg tttgtcgtct cagaatgtcg ccatgatctt
     3301 cagtaagcgt agcactcgta ctcgtgtatc agtcgaaagt gctgtcagtt gccttggcgg
     3361 caatgccatg tttttaggta aagatgatat ccaattagga gtgaatgagt ctttatacga
     3421 tacttcaaaa gttatatcaa gtatggtttc tggtattgtt gctcgcgtaa acaaatattc
     3481 agatgttgct acgctcgcta aacatgcctc ttgtcctgtt attaatggtc tttgtgacac
     3541 gtttcatcct ctccaggcgt tagcagattt actcactatt aaggaaactt tcaagagctt
     3601 tgacggttta aaagttgcat gggtaggtga tgccaataac gttttacatg atttgatgat
     3661 tgcaaatgcc aaggttggta ttcatactag cgttgctaag cccaaggacg tcaacgttcg
     3721 cgacgatatt ttatcaatcg ttaacgaagc tgcgaacgaa aacggttcta cttttgaaat
     3781 tgttaacgac cccaaggttg ctgttaaaaa tgctgatatc gtcgttaccg acacttggat
     3841 ttctatgggt caagaagctg agaaggaaca acgtcttaaa caatttacag gattccaagt
     3901 caccggagag atcatgaaac ttgccaaacc tagctgcaaa ttcatgcatt gtttaccccg
     3961 tcatcccgag gaagtgtctg acgaggtatt ttatggagag aacagtttag ttttccaaga
     4021 agcagagaac cgcaaatgga ccactgttgc tgtcttggag gctttgcttg tcaaccgtgg
     4081 tgaaattctt cctcctgcct cagcctaaac tgaaatcctt aatttcattt ttatctgggg
     4141 aatgtgcaaa caatgccaac tttataattt tttttattgg gcgtgcaaat tgtgatattc
     4201 atttttgaaa ttttatgtac tttgtttcac aaaaagaaaa ttgacttttg agaaaatttt
     4261 ttaattgaca attacttttc agatttttta tgcgcaggac acaggcttct aaagacgtac
     4321 ttatacgtta aaaaagggga tggatcaatt tggctcatta gtccaacaag gtttgtgtaa
     4381 aattattacc cgcttaatgc tatcctctta actagatatt tcaaggtatc aagcaatcat
     4441 agtattaagc actattatat gcaaaataca atttaaaatt aatagaaact acaaataagc
     4501 gcaacactca taaggaaaaa ggtttgggac aataaaggtt tcgtcaatgc ggaatctacg
     4561 acagaaagaa aacttagtgt aggcatcgaa atttattata tttcaagcat gtatgctagt
     4621 aaaataaaac gtaaactcat tgattcaaga aatctataga aaaaaagcta gcgtgacgca
     4681 gacattcgaa tggcatgccc tacaacaact aagaaaatgg ctatcatgcg gaagctggtg
     4741 agaagaacag catcgggaca agggaaggaa gaacaaagac aaagaaaaca aaagaaagca
     4801 attgaaaaca aaacaaaaca attttcattc cttctcttat cattcctttt cttttctttt
     4861 ctctcattca acgcactcca tcgtatccgt attcctctta ttttttctct ttctctatat
     4921 ccatttcttt ctctctaggt gtgtcctctc tctctcttca atttctctac tccgcattcc
     4981 aacgcatcct tcccccaacc tcccatttcc tccttacggc ccgatagcga tcgtctttcc
     5041 ctcgctatca ctcgctaccg gcccctcctc tgcaccgtaa cctcctacgt atttaccata
     5101 tcataaagtt ttttccgacg cttatcgctg accccctgtc gccctcctat tggcttccgg
     5161 attatcttct tgtccataag gtgatccatg cttcctgaag attcccgaaa tgtgtccact
     5221 ttggcgggga atcattccat ccacttcttt ctctctcgct ttcctcattc ggcgctcccc
     5281 ttccgcgtct cattggtctt ccgctccgtt tttgctttgc cgatgttact tggggagagg
     5341 tgcgataatc ctttcgcaaa aactcggttt gacgcctccc atggtataaa tagtgggtgg
     5401 tggacaggtg ccttcgcttt tctttaagca agagaattcc attgtcttga ctatcacaaa
     5461 cttttaagtc ttttcttttt tctaaccaca tacatatgac ttattttccg gaagaggtgg
     5521 tcgaacacat tttttctttc ttgccagcgc agcgagacag aaataccgtt tctttggttt
     5581 gtaaagtttg gtatgaaatc gaaagactgt ccaggcgagg cgtgtttgtc ggtaactgct
     5641 atgcggtcag agctggaagg gtggcagcga gattcccgaa tgtgagggcg ttgactgtta
     5701 agggaaaacc acattttgcg gactttaact tggttcctcc tgactgggga ggctacgccg
     5761 ggccctggat tgaggcggca gctcgaggtt gtcatggctt ggaggagctc cgcatgaaac
     5821 gcatggtggt ctccgacgag tctttggagt tgctcgctcg gagcttccct aggttccggg
     5881 ctttggtcct gatatcatgt gaaggattta gtacggacgg attggcagcg gtagccagtc
     5941 attgcaaact tctgagggag ttggacttgc aagagaacga ggtggaggat cgaggacctc
     6001 gctggctgtc atgctttcct gacagttgca catcactggt ttcactcaat ttcgcctgta
     6061 ttaagggaga agtgaatgca ggttccctcg aacgactcgt ttcccggagc cccaatctcc
     6121 gatccttgag actcaaccga agtgtcagtg tggacacgct tgccaagatc ctcttgagaa
     6181 cccctaatct cgaggatctc ggaaccggca accttactga tgatttccaa acagagagct
     6241 attttaaact cacctcagcg ctcgaaaaat gcaaaatgtt gaggtctttg agcgggtttt
     6301 gggatgcatc cccagtctgt ctcagcttca tatatccgtt gtgcgctcaa ctcacagggt
     6361 tgaatctttc ttatgcacct accctggacg cttccgattt gaccaagatg atctctcgct
     6421 gcgttaagtt gcaacgcttg tgggtcctgg attgcataag tgacaagggg ctgcaagtgg
     6481 tggcttccag ttgtaaagac ctccaggaac tgcgggtatt tcctagcgat ttctatgtag
     6541 cgggatatag cgctgtcact gaagaaggcc ttgtggcagt ctctcttgga tgcccgaagt
     6601 tgaactcatt gctgtacttc tgccaccaaa tgacaaacgc agctcttgtt accgttgcta
     6661 agaactgtcc caatttcacg cggttccgcc tttgcatttt ggagcctggg aagccggacg
     6721 tggtcacatc acaacctttg gacgaggggt tcggcgcgat agtgagggaa tgtaaaggcc
     6781 tccaaagact tagtatcagt ggtctgctta ccgacaaggt cttcatgtat ataggaaaat
     6841 acgcaaagca gctcgaaatg ttgtctatcg cattcgcagg tgattccgac aagggtatga
     6901 tgcatgtaat gaatgggtgt aagaacctca ggaaactgga gatacgagac tcacctttcg
     6961 gtgacgctgc cctccttggt aactttgcgc ggtatgagac gatgcggtca ctctggatgt
     7021 caagttgcaa cgttacgctg aaaggctgtc aagttcttgc atcaaagatg ccaatgctta
     7081 atgttgaagt tataaatgaa agagacggca gcaatgaaat ggaggaaaat cacggggact
     7141 tgcccaaggt tgaaaaattg tatgtctata gaaccacggc gggtgccaga gatgacgcgc
     7201 ccaatttcgt gaaaatcctt ggatcaggtc ctaaaaagaa gagaaaggtc taagtcgaca
     7261 cttctaaata agcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa
     7321 aaataagtgt atacaaattt taaagtgact cttaggtttt aaaacgaaaa ttcttattct
     7381 tgagtaactc tttcctgtag gtcaggttgc tttctcaggt atagtatgag gtcgctctta
     7441 ttgaccacac ctctaccggc agatccgcta gggataacag ggtaatatac tagtgatctg
     7501 ggcaagtaga gtgtgcatca tcgaaatccg gcttctttca taaacttatc gtattgctct
     7561 ttaagtgctt tgcttttatc aataaatggc tttttcttgt cttccgccaa ttccttccac
     7621 gcctttgatg cattttgggt ctcttcaacg agtgtggaaa tggcttctgg aatacctaac
     7681 tcttgacgta gcttgggatt gtttcgaagc tccttgtaaa agaggataaa agcagaagaa
     7741 ggttttttag gaacatctgg atgacgaagt ctgtaatagc gactgcccgt gctgtgagcc
     7801 tccttcgagc gacgacgatt ttcactatcg atttctgtag gtgtaaagga tcgtagagtc
     7861 tccataaaac gctttttctc agcatcgaac tgctcctgga gtttgttttt atacttttgg
     7921 atctcatcgg cagtcaaagc tgcggccgca agcttggcgt aatcatggtc atagctgttt
     7981 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     8041 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     8101 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     8161 gggagaggcg gtttgcgtat tgggcgc
//
