LOCUS       Exported                3996 bp ds-DNA     circular SYN 04-FEB-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3996)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Feb 4, 2020 from SnapGene Viewer 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3996
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      403..412
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             982..1002
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1011..1028
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1057..1281
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1352..1368)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1376..1392
                     /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(1400..1430)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1445..1466
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1754..2342)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2513..3373)
                     /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(3374..3478)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(3748..3996,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat gggcaagatc
      421 gacaccaccg tgaccgaggt gaaggagaac ggcaagacct acctgagact gctgaagggc
      481 aacgagcagc tgaaggccgt gagcgacaag gccgtggccg gcgtgaacct gttccccggc
      541 gccaagatcg gcagcttcct ggtgagacag gacaacatcg tggtgttccc cgacaacaag
      601 ggcgagttcg acctggactt cttcaacctg ctgaacgaca acttcgagac cctggtggag
      661 tacgccaaga tggccgactg cctggacatc gccttcgaca tcaacgagaa gagctacttc
      721 aacatgatca tgtggctgat gaagaacatc gacgagaact ggagccagag cccctacggc
      781 gagagcttct acagcagcaa ggacatcgac tggggctaca agcccgaggg cagcctgaga
      841 gtgagcgacc actggaactt cggccaggac ggcgagcact gccccaccgc cgagcccgtg
      901 gacggctggg ccgtgtgcaa gttcgagaac ggcaagtacc acctgatcaa gaagttcgga
      961 tccggcggtg gaggcagtgg gcccaagaag aagaggaaag tctgaccggt catcatcacc
     1021 atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     1081 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     1141 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     1201 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     1261 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tcgataccgt
     1321 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1381 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctagggtg
     1441 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1501 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1561 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1621 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1681 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1741 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1801 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1861 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1921 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     1981 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2041 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2101 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2161 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     2221 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2281 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2341 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2401 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2461 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2521 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2581 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     2641 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     2701 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     2761 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     2821 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     2881 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     2941 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3001 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3061 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3121 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3181 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3241 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3301 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3361 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3421 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3481 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcgatct cccgatcccc
     3541 tagggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     3601 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     3661 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
     3721 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
     3781 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     3841 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     3901 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gactatttac
     3961 ggtaaactgc ccacttggca gtacatcaag tgtatc
//
