LOCUS       V039736                 5855 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039736
VERSION     V039736
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..1432
                     /label="ARF6(NM_001663)"
                     /note="ARF6(NM_001663)"
                     /gene="ARF6"
     CDS             1433..1456
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             1457..1570
                     /codon_start=1
                     /note="selected from a peptide library; binds streptavidin
                     with nanomolar affinity (Keefe et al., 2001)"
                     /product="streptavidin-binding peptide"
                     /transl_table=1
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
                     /label="SBP"
     polyA_signal    1617..1841
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      1887..2315
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2329..2658
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2725..3324
                     /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    3498..3619
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(3668..3684)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3692..3708
                     /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(3716..3746)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    3761..3782
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4070..4655)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(4826..5686)
                     /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(5687..5791)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gggaaggtgc tatccaaaat cttcgggaac aaggaaatgc ggatcctcat
      961 gttgggcctg gacgcggccg gcaagacaac aatcctgtac aagttgaagc tgggccagtc
     1021 ggtgaccacc attcccactg tgggtttcaa cgtggagacg gtgacttaca aaaatgtcaa
     1081 gttcaacgta tgggatgtgg gcggccagga caagatccgg ccgctctggc ggcattacta
     1141 cactgggacc caaggtctca tcttcgtagt ggactgcgcc gaccgcgacc gcatcgatga
     1201 ggctcgccag gagctgcacc gcattatcaa tgaccgggag atgagggacg ccataatcct
     1261 catcttcgcc aacaagcagg acctgcccga tgccatgaaa ccccacgaga tccaggagaa
     1321 actgggcctg acccggattc gggacaggaa ctggtatgtg cagccctcct gtgccacctc
     1381 aggggacgga ctctatgagg ggctcacatg gttaacctct aactacaaat ctgattacaa
     1441 ggatgacgac gataagatgg acgagaagac caccggctgg cggggcggcc acgtggtgga
     1501 gggcctggcc ggcgagctgg agcagctgcg ggccaggctg gagcaccacc ctcagggcca
     1561 gcgggagccc tgactcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     1621 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     1681 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     1741 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     1801 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     1861 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     1921 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     1981 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2041 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2101 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     2161 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     2221 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     2281 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     2341 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2401 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2461 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     2521 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     2581 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     2641 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     2701 gagacaggat gaggatcgtt tcgcatgacc gagtacaagc ccacggtgcg cctcgccacc
     2761 cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     2821 acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     2881 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     2941 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     3001 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     3061 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     3121 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     3181 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     3241 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     3301 atgacccgca agcccggtgc ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     3361 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     3421 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     3481 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3541 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3601 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     3661 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3721 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     3781 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3841 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3901 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3961 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4021 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4081 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4141 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4201 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4261 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4321 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4381 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4441 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4501 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4561 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     4621 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4681 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4741 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4801 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4861 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4921 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4981 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5041 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5101 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5161 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5221 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5281 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5341 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5401 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5461 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5521 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5581 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5641 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5701 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5761 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5821 tcgacggatc gggagatctc ccgatcccct atggt
//
