LOCUS       pAWOSUMO3C-14M_z        6599 bp DNA     circular SYN 28-MAR-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6599)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 28, 2023 from SnapGene 6.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..131
                     /gene="bla"
                     /label=AmpR promoter
     CDS             132..992
                     /codon_start=1
                     /label=Amp(R)
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1147..1766
                     /label=pBR322_origin
     misc_feature    1937..2076
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2162..2184
                     /label=pGEX_3_primer
     CDS             complement(2178..2369)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    complement(3681..4772)
                     /label=lacI
     promoter        complement(4764..4841)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5047..5066
                     /label=pBRrevBam_primer
     promoter        5150..5168
                     /label=T7_promoter
     protein_bind    5169..5193
                     /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)."
     RBS             5208..5230
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5237..5239
                     /codon_start=1
                     /label=Start
                     /translation="M"
     CDS             5246..5263
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5270..5557
                     /codon_start=1
                     /label=Smt3
                     /translation="DSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRR
                     LMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             5558..5581
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5585..6340
                     /codon_start=1
                     /label=14M_zTDP2
                     /note="CI:V1=NdeI,V2=EcoRI,I1=NdeI,I2=EcoRI"
                     /translation="MDSKLSIISWNVDGLDTNNLSDRARGLCSYLALYTPDVVFLQELI
                     PAYVQYLKKRAVSYLFFEGSDDGYFTGIMLRKSRVKFLESEIICFPTTQMMRNLLIAQV
                     TFSGQKLYLMTSHLESTRNQSQERTKQLRVVLQKIKEAPEDAIVIFAGDTNLRDAEVAN
                     VGGLPAGVCDVWEQLGKQEHCRYTWDTQANSNLGITAACKLRFDRIFLRSAKTAPPVTP
                     DHMALIGMEKLDCGRYTSDHWGIYCTFNT"
     primer_bind     complement(6434..6452)
                     /label=T7_Terminal_primer
     terminator      6448..6495
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
       61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
      181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
      241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
      301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
      361 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
      421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
      481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
      541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
      601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
      661 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
      721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
      781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
      841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
      901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
      961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     1081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     1321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     1801 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     1861 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     1921 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     1981 accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     2041 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     2101 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     2161 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     2221 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     2281 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     2341 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
     2401 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
     2461 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
     2521 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
     2581 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
     2641 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
     2701 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
     2761 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
     2821 atcatgcgca cccgtggcca ggacccaacg ctgcccgaga tgcgccgcgt gcggctgctg
     2881 gagatggcgg acgcgatgga tatgttctgc caagggttgg tttgcgcatt cacagttctc
     2941 cgcaagaatt gattggctcc aattcttgga gtggtgaatc cgttagcgag gtgccgccgg
     3001 cttccattca ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga
     3061 caaggtatag ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg
     3121 cataaatcgc cgtgacgatc agcggtccaa tgatcgaagt taggctggta agagccgcga
     3181 gcgatccttg aagctgtccc tgatggtcgt catctacctg cctggacagc atggcctgca
     3241 acgcgggcat cccgatgccg ccggaagcga gaagaatcat aatggggaag gccatccagc
     3301 ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa
     3361 tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg
     3421 cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc
     3481 ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa
     3541 agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga
     3601 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     3661 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3721 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg
     3781 gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga
     3841 gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg
     3901 gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag
     3961 atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc
     4021 tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt
     4081 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
     4141 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
     4201 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
     4261 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
     4321 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
     4381 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
     4441 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
     4501 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
     4561 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
     4621 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
     4681 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
     4741 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
     4801 catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc
     4861 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
     4921 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     4981 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     5041 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     5101 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc
     5161 actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt
     5221 aagaaggaga tataccatgg gcagccatca ccatcaccat cacggcagcg acagcgaagt
     5281 caatcaggaa gcaaaaccgg aagttaaacc ggaagtcaaa ccggaaaccc atattaacct
     5341 gaaagtctca gatggcagct ctgaaatttt ctttaaaatt aagaaaacca cgccgctgcg
     5401 tcgcctgatg gaagcatttg ctaaacgtca gggcaaagaa atggattcgc tgcgtttcct
     5461 gtatgacggt attcgcatcc aggcagatca aaccccggaa gatctggaca tggaagataa
     5521 cgacattatc gaagctcacc gtgaacagat tggcggtctg gaagttctgt tccaggggcc
     5581 ccatatggat agcaaactga gcattattag ctggaatgtt gatggcctgg ataccaataa
     5641 tctgagcgat cgtgcacgtg gtctgtgtag ctatctggca ctgtatacac cggatgttgt
     5701 ttttctgcaa gaactgattc ctgcctatgt gcagtatctg aaaaaacgtg cagttagcta
     5761 cctgtttttt gaaggtagtg atgatggtta tttcaccggt attatgctgc gtaaaagccg
     5821 tgttaaattt ctggaaagcg aaatcatttg ttttccgacc acacagatga tgcgtaatct
     5881 gctgattgca caggttacct ttagcggtca gaaactgtat ctgatgacca gccatctgga
     5941 aagtacccgt aatcagagcc aagaacgtac caaacagctg cgtgttgtgc tgcagaaaat
     6001 caaagaagca cctgaggacg ccattgttat ttttgccggt gataccaatc tgcgtgatgc
     6061 cgaagttgca aatgttggtg gtctgcctgc cggtgtttgt gatgtttggg aacagctggg
     6121 taaacaagaa cattgtcgtt atacctggga tacccaggca aatagcaatc tgggtattac
     6181 cgcagcatgt aaactgcgtt ttgatcgtat ttttctgcgt agcgcaaaaa ccgcaccgcc
     6241 tgtgacaccg gatcacatgg cactgattgg tatggaaaaa ctggattgtg gtcgctatac
     6301 cagcgatcat tggggtattt attgtacctt taacacctaa taagaattct gcagatatcc
     6361 atcacactgg cggccgctcg agcagatccg gctgctaaca aagcccgaaa ggaagctgag
     6421 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     6481 ttgaggggtt ttttgctgaa aggaggaact atatccggat aattcttgaa gacgaaaggg
     6541 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgt
//
