LOCUS       Exported                6451 bp ds-DNA     circular SYN 27-OCT-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6451)
  AUTHORS   Kazuhiko
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Dec 20, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6451
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    30..180
                     /label=Tolmini-L
     promoter        187..205
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    323..457
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        462..479
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(523..1224)
                     /codon_start=1
                     /label=zTagRFP658
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTQRIKV
                     VEGGPLPFAFDILATSFMYGSHTFIDHTQGIPDFWKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLHDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEAHTEMLYPTDGGLEGRTALALK
                     LVGGGHLICNFKTIYRSKKPAKNLKMPGVYYVDYRLERIEEADNETYVEQHEVAVARYC
                     DLPTKLGHKLN"
     misc_feature    1275..1402
                     /label=CpG free intron
     promoter        complement(1433..1657)
                     
     misc_feature    1658..2083
                     /label=mouse CpG free CMV enh
     misc_feature    2219..2641
                     /label=mouse CpG free CMV enh
     promoter        2642..2866
                     /label=human CpG free EF1 promoter
     misc_feature    2897..3024
                     /label=CpG free synthetic intron
     CDS             3058..3780
                     /codon_start=1
                     /label=mClover3
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK*"
     promoter        complement(3789..3806)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3811..3945
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(4048..4066)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    4073..4273
                     /label=Tol2miniR
     misc_feature    complement(4075..4095)
                     /label=934
     primer_bind     complement(4302..4318)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4326..4342
                     /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(4350..4380)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4395..4416
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4704..5292)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5463..6323)
                     /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(6324..6428)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaatc ccatatgggc agaggtgtaa aaagtactca aaaattttac
       61 tcaagtgaaa gtacaagtac ttagggaaaa ttttactcaa ttaaaagtaa aagtatctgg
      121 ctagaatctt acttgagtaa aagtaaaaaa gtactccatt aaaattgtac ttgagtatta
      181 gatctgtaat acgactcact atagggacat agaatgcagg taccgggccc aatgcattgg
      241 cgccgcggcc ggccgcgaat taaaaaacct cccacacctc cccctgaacc tgaaacataa
      301 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
      361 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
      421 gtccaaactc atcaatgtat cttatcatgt ctggatctac gtaatacgac tcactatagt
      481 tctagcctcg agaaggccgg ccataagatc taatggcgcg cctcagttca gcttgtgtcc
      541 cagctttgta ggcaggtcac agtatctagc cacagccacc tcgtgctgct ccacgtatgt
      601 ctcgttgtca gcctcctcga ttctctccag tctgtagtcc acgtagtaca ctccaggcat
      661 cttcaggttc ttagcaggct tcttgcttct gtagattgtc ttgaagttac agatcaggtg
      721 tcctcctccc accagcttca gagccagagc tgttcttccc tccagtcctc cgtctgtagg
      781 gtacagcatc tctgtgtgag cctcccatcc cagtgtcttc ttctgcatca caggtccgtt
      841 gctagggaag ttcactcctc tgatcttcac gttgtagatc agacatccgt cgtgcaggct
      901 tgtgtcctgt gtagctgtca gcactcctcc gtcctcgtat gttgtcactc tctcccatgt
      961 gaatccctca gggaagctct gcttccagaa gtcagggatt ccctgtgtgt ggtcgatgaa
     1021 tgtgtggctt ccgtacatga agcttgtagc caggatgtcg aaagcgaaag gcagaggtcc
     1081 tccctccacc accttgattc tctgtgtctg tgttccctcg taaggctttc cctctccctc
     1141 gcttgtacac ttgaagtggt ggttgttcac tgttccctcc atgtacagct tcatgtgcat
     1201 gttctcctta atcagctcgc tcatacccat ggtggatatc gtttaaacaa ttctggaagc
     1261 tactgtacac caacctgtca ggagaggaaa gagaagaagg ttagtacaat tgtctagggc
     1321 tgcagggttc acagtgccac ttttcctgca ctgccccacc tcctgcccac cctttcccag
     1381 gcatagacag tcagtgactt accaaactca caggagggag aaggcagaag cttgaatgtt
     1441 cacagagact actgcactta tatatggttc tcccccaccc tggggaaaaa ggtggagcca
     1501 gtacaccaca tcactttccc agtttaccca agccccacct tctctaggca ccagttcaat
     1561 tgcccacccc tccccccaac ttctcaggga ctgtgggcca tgtgctctct gcccactgag
     1621 gggcactcag ccctcaagca tgctcttctc cactagtcac ccctattgac cttatgtatg
     1681 tgccaataat gggaaaaacc cattgacdss tcacccccta ttgacctttt gtactgggca
     1741 aaacccaatg gaaagtccct attgactcag tgtacttggc tccaatggga ctttcctgtt
     1801 gattcacccc tattgacctt atgtactggg caaaacccat tggaaagtcc ctaatgactc
     1861 agtatacatg tcagtaatgg gaaaaaccca ttggcttacc tcccattgac cttatgtact
     1921 gggcaaaacc cattggaaag tccctattga ctcaatgtac ttggctccaa tgggactttc
     1981 ctgttgactc accccctatt gaccttatgt actgggcaaa acccaatgga aagtccctat
     2041 tgagtcagtg tacttggctc caatgggttt ttcccattga ctcctgcatt aatcgattta
     2101 aatgctaggg ctgctctcca atctctgtca tggcagggcc actggcatcc ctgaagttgg
     2161 tggagaccac ctcactccac tcagcataca gctcatccag tcctcttaat taatgcagga
     2221 gtcaatggga aaaacccatt ggagccaagt acactgactc aatagggact ttccattggg
     2281 ttttgcccag tacataaggt caataggggg tgagtcaaca ggaaagtccc attggagcca
     2341 agtacattga gtcaataggg actttccaat gggttttgcc cagtacataa ggtcaatggg
     2401 aggtaagcca atgggttttt cccattactg acatgtatac tgagtcatta gggactttcc
     2461 aatgggtttt gcccagtaca taaggtcaat aggggtgaat caacaggaaa gtcccattgg
     2521 agccaagtac actgagtcaa tagggacttt ccattgggtt ttgcccagta caaaaggtca
     2581 atagggggtg agtcaatggg tttttcccat tattggcaca tacataaggt caataggggt
     2641 gactagtgga gaagagcatg cttgagggct gagtgcccct cagtgggcag agagcacatg
     2701 gcccacagtc cctgagaagt tggggggagg ggtgggcaat tgaactggtg cctagagaag
     2761 gtggggcttg ggtaaactgg gaaagtgatg tggtgtactg gctccacctt tttccccagg
     2821 gtgggggaga accatatata agtgcagtag tctctgtgaa cattcaagct tctgccttct
     2881 ccctcctgtg agtttggtaa gtcactgact gtctatgcct gggaaagggt gggcaggagg
     2941 tggggcagtg caggaaaagt ggcactgtga accctgcagc cctagacaat tgtactaacc
     3001 ttcttctctt tcctctcctg acaggttggt gtacagtagc ttccagaatt cgcaaacatg
     3061 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     3121 gacgtaaacg gccacaagtt cagcgtccgc ggcgagggcg agggcgatgc caccaacggc
     3181 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     3241 gtgaccacct tcggctacgg cgtggcctgc ttcagccgct accccgacca catgaagcag
     3301 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catctctttc
     3361 aaggacgacg gcacctacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     3421 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     3481 ctggagtaca acttcaacag ccactacgtc tatatcacgg ccgacaagca gaagaactgc
     3541 atcaaggcta acttcaagat ccgccacaac gttgaggacg gcagcgtgca gctcgccgac
     3601 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     3661 ctgagccatc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     3721 ctggagttcg tgaccgccgc cgggattaca catggcatgg acgagctgta caagtaataa
     3781 atctagaact atagtgagtc gtattacgta gatccagaca tgataagata cattgatgag
     3841 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     3901 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     3961 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaatt cgcggccgcc
     4021 accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgggc ccaagtgatc
     4081 tccaaaaaat aagtactttt tgactgtaaa taaaattgta aggagtaaaa agtacttttt
     4141 tttctaaaaa aatgtaatta agtaaaagta aaagtattga tttttaattg tactcaagta
     4201 aagtaaaaat ccccaaaaat aatacttaag tacagtaatc aagtaaaatt actcaagtac
     4261 tttacacctc tgggtactgt ttcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     4321 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4381 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4441 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4501 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4561 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4621 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4681 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4741 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4801 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4861 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4921 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4981 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5041 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5101 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     5161 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5221 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5281 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5341 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5401 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5461 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5521 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5581 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5641 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5701 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5761 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5821 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5881 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5941 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6001 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6061 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6121 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6181 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6241 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6301 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6361 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6421 gttccgcgca catttccccg aaaagtgcca c
//
