LOCUS       V034709                 7054 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034709
VERSION     V034709
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             925..2805
                     /label="CHRNA4(NM_000744)"
                     /note="CHRNA4(NM_000744)"
                     /gene="CHRNA4"
     CDS             2818..2883
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2918..3142
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3188..3616
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3630..3959
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3971..4570
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    4730..4851
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4900..4916)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4924..4940
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(4948..4978)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4993..5014
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5302..5887)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6058..6918)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(6919..7023)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 aagctttcta gagaattcgc caccatggag ctagggggcc ccggagcgcc gcggctgctg
      961 ccgccgctgc tgctgcttct ggggaccggc ctcctgcgcg ccagcagcca tgtggagacc
     1021 cgggcccacg ccgaggagcg gctcctgaag aaactcttct ccggttacaa caagtggtcc
     1081 cgacccgtgg ccaacatctc ggacgtggtc ctcgtccgct tcggcctgtc catcgctcag
     1141 ctcattgacg tggatgagaa gaaccagatg atgaccacga acgtatgggt gaagcaggag
     1201 tggcacgact acaagctgcg ctgggaccca gctgactatg agaatgtcac ctccatccgc
     1261 atcccctccg agctcatctg gcggccggac atcgtcctct acaacaatgc tgacggggac
     1321 ttcgcggtca cccacctgac caaggcccac ctgttccatg acgggcgggt gcagtggact
     1381 cccccggcca tttacaagag ctcctgcagc atcgacgtca ccttcttccc cttcgaccag
     1441 cagaactgca ccatgaaatt cggctcctgg acctacgaca aggccaagat cgacctggtg
     1501 aacatgcaca gccgcgtgga ccagctggac ttctgggaga gtggcgagtg ggtcatcgtg
     1561 gatgccgtgg gcacctacaa caccaggaag tacgagtgct gtgccgagat ctacccggac
     1621 atcacctatg ccttcgtcat ccggcggctg ccgctcttct acaccatcaa cctcatcatc
     1681 ccctgcctgc tcatctcctg cctcaccgtg ctggtcttct acctgccctc cgagtgtggc
     1741 gagaagatca cgctgtgcat ctccgtgctg ctgtcgctca ccgtcttcct gctgctcatc
     1801 accgagatca tcccgtccac ctcactggtc atcccactca tcggcgagta cctgctgttc
     1861 accatgatct tcgtcaccct gtccatcgtc atcacggtct tcgtgctcaa cgtgcaccac
     1921 cgctcgccac gcacgcacac catgcccacc tgggtacgca gggtcttcct ggacatcgtg
     1981 ccacgcctgc tcctcatgaa gcggccgtcc gtggtcaagg acaattgccg gcggctcatc
     2041 gagtccatgc ataagatggc cagtgccccg cgcttctggc ccgagccaga aggggagccc
     2101 cctgccacga gcggcaccca gagcctgcac ccgccctcac cgtccttctg tgtccccctg
     2161 gatgtgccgg ctgagcctgg gccttcctgc aagtcaccct ccgaccagct ccctcctcag
     2221 cagcccctgg aagctgagaa agccagcccc cacccctcgc ctggaccctg ccgcccgccc
     2281 cacggcaccc aggcaccagg gctggccaaa gccaggtccc tcagcgtcca gcacatgtcc
     2341 agccctggcg aagcggtgga aggcggcgtc cggtgccggt ctcggagcat ccagtactgt
     2401 gttccccgag acgatgccgc ccccgaggca gatggccagg ctgccggcgc cctggcctct
     2461 cgcaacaccc actcggctga gctcccaccc ccagaccagc cctctccgtg caaatgcaca
     2521 tgcaagaagg agccctcttc ggtgtccccg agcgccacgg tcaagacccg cagcaccaaa
     2581 gcgccgcccc cgcacctgcc cctgtcgccg gccctgaccc gggcggtgga gggcgtccag
     2641 tacattgcag accacctgaa ggccgaagac acagacttct cggtgaagga ggactggaag
     2701 tacgtggcca tggtcatcga ccgcatcttc ctctggatgt tcatcatcgt ctgcctgctg
     2761 gggacggtgg gcctcttcct gccgccctgg ctggctggca tgatcctcga gaccggtgac
     2821 tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac
     2881 aagtgaccgg taagtttaaa ccgctgatca gcctcgactg tgccttctag ttgccagcca
     2941 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3001 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3061 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3121 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     3181 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3241 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3301 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3361 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3421 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3481 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3541 gatttataag ggattttggc catttcggcc tattggttaa aaaatgagct gatttaacaa
     3601 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3661 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3721 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3781 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3841 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3901 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3961 agctcccggg atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     4021 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     4081 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     4141 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     4201 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     4261 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     4321 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     4381 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     4441 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     4501 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     4561 cggtgcctga ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4621 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4681 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4741 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4801 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4861 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4921 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4981 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5041 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5101 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5161 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5221 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5281 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5341 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5401 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5461 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5521 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5581 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5641 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5701 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5761 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5821 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5881 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5941 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6001 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6061 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6121 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6181 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6241 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6301 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6361 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6421 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6481 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6541 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6601 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6661 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6721 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6781 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6841 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6901 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6961 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7021 gcgcacattt ccccgaaaag tgccacctga cgtc
//
