LOCUS       Exported                5059 bp DNA     circular SYN 05-MAY-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5059)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 5, 2022 from SnapGene Viewer 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5059
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     terminator      82..168
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(180..892)
                     /codon_start=1
                     /label=Ci_mutant_bagh
                     /translation="MAQKRNH*HKSSLRTHVALKQFMKKRKMNLAYPRNLSQTRWGWGS
                     QALVLYLMASMH*MLITPHCLQKFSKLALKNLALQSPEKSTRCMKRLVCSRHLEVSMST
                     LFFLMFRQGCSHLSLEPLPKVMRRDG*AQPKKPVILHSGLRLKVIP*PHQQAPSQAFLT
                     EC*FSLTLSRLLSQVISA*PDLGVMSLPSRN*SGIAVRCFYNH*THSTQ*SHAMRVVPL
                     WGKLSLVSGLKRRLA"
     protein_bind    950..968
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
     protein_bind    975..993
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
     protein_bind    1077..1095
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
     protein_bind    1102..1118
                     /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)."
     protein_bind    1133..1151
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
     protein_bind    1157..1173
                     /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)."
     protein_bind    1181..1197
                     /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             1209..1220
     CDS             1227..1946
                     /codon_start=1
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      1958..2044
                     /label=rrnB T1 terminator
     terminator      2159..2245
                     /label=rrnB T1 terminator
     CDS             complement(2257..2976)
                     /codon_start=1
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     RBS             2983..2997
                     /label=700
     promoter        3004..3058
                     /label=Pr
     rep_origin      complement(3264..3852)
                     /direction=LEFT
                     /label=ori
     terminator      3940..4034
                     /label=lambda t0 terminator
     CDS             complement(4064..4924)
                     /codon_start=1
                     /label=AmpR
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4925..5029)
                     /label=AmpR promoter
ORIGIN
        1 ctactgggac gaagacgaac agtcgacaac gaagacgcca acatcaagat ctcgcatttg
       61 ttcctaggtc tagggcggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
      121 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctctagat
      181 cagccaaacg tctcttcagg ccactgacta gcgataactt tccccacaac ggaacaactc
      241 tcattgcatg ggatcattgg gtactgtggg tttagtggtt gtaaaaacac ctgaccgcta
      301 tccctgatca gtttcttgaa ggtaaactca tcacccccaa gtctggctat gcagaaatca
      361 cctggctcaa cagcctgctc agggtcaacg agaattaaca ttccgtcagg aaagcttggc
      421 ttggagcctg ttggtgcggt catggaatta ccttcaacct caagccagaa tgcagaatca
      481 ctggcttttt tggttgtgct tacccatctc tccgcatcac ctttggtaaa ggttctaagc
      541 tcaggtgaga acatccctgc ctgaacatga gaaaaaacag ggtactcata ctcacttcta
      601 agtgacggct gcatactaac cgcttcatac atctcgtaga tttctctggc gattgaaggg
      661 ctaaattctt caacgctaac tttgagaatt tttgcaagca atgcggcgtt ataagcattt
      721 aatgcattga tgccattaaa taaagcacca acgcctgact gccccatccc catcttgtct
      781 gcgacagatt cctgggataa gccaagttca tttttctttt tttcataaat tgctttaagg
      841 cgacgtgcgt cctcaagctg ctcttgtgtt aatggtttct tttttgtgcc atggtacctt
      901 tctcctcttt aatgaattcg gtcagtgcgt cctgctgatg tgctcagtat ctctatcact
      961 gatagggatg tcaatctcta tcactgatag ggactcgagt taccaggatc tttattatca
     1021 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcac ctcgagtccc
     1081 tatcagtgat agagattgac attgtgagcg gataacaaga tactgagcac cctccctatc
     1141 agtgatagag agataattgt gagcggataa caattgataa ttgtgagcgg ataacaattg
     1201 aattcattaa agaggagaaa ggtaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     1261 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     1321 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     1381 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     1441 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1501 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1561 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1621 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     1681 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     1741 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1801 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     1861 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1921 tcggcatgga cgagctgtac aagtaatcta gaggcatcaa ataaaacgaa aggctcagtc
     1981 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     2041 aaatccgccg ccctagacct aggcgatgac ggaaaaagag atcgtcttaa tgtatagatt
     2101 ttcgtggatt acgtcgccag tcaggatctc gcatttgttc ctaggtctag ggcggcggat
     2161 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     2221 tcgactgagc ctttcgtttt atttgatgcc tctagattac ttgtacagct cgtccatgcc
     2281 gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt
     2341 ggggtctttg ctcagggcgg actgggtgct caggtagtgg ttgtcgggca gcagcacggg
     2401 gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc tgccgtcctc
     2461 gatgttgtgg cggatcttga agttcacctt gatgccgttc ttctgcttgt cggccatgat
     2521 atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc
     2581 cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac
     2641 ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta
     2701 gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct
     2761 gaagcactgc acgccgtagg tcagggtggt cacgagggtg ggccagggca cgggcagctt
     2821 gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc cctcgccctc
     2881 gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac
     2941 caccccggtg aacagctcct cgcccttgct caccatggta cctttctcgt ctgctgtgaa
     3001 ttcgtacatg caaccattat caccgccaga ggtaaaatag tcaacacgca cggtgttact
     3061 cgagttacca ggatcctggc agaagctatt cattctgcag ttccgtgctc caaaacaacc
     3121 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3181 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3241 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3301 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3361 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3421 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     3481 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3541 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3601 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3661 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     3721 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3781 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3841 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3901 aactcacgtt aagggatttt ggtcatgact agtgcttgga ttctcaccaa taaaaaacgc
     3961 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc
     4021 tatcaacagg agtccaagcg agctcgtaaa cttggtctga cagttaccaa tgcttaatca
     4081 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4141 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4201 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4261 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4321 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4381 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4441 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4501 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4561 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4621 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4681 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4741 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4801 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4861 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4921 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4981 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5041 gaaaagtgcc acctgacgt
//
