LOCUS       V032685                 6594 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032685
VERSION     V032685
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..1274
                     /label="Ssr4(NM_001166480)"
                     /note="Ssr4(NM_001166480)"
                     /gene="Ssr4"
     CDS             1281..1346
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1370..1958
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2024..2072
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2274..2702
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2716..3045
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3112..3906
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    4082..4203
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4252..4268)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4276..4292
                     /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(4300..4330)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4345..4366
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4654..5242)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5413..6273)
                     /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(6274..6378)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc ggcgatggca tctttcggcg
      781 ccctggcact actcctgctg tctggcctat cttgctgctc agcagaggcc tgcctggaac
      841 cacagatcac cccttcttac tatacaactt cagatgccgt catttctaca gagactgtat
      901 tcatcgtgga gatttcactg acctgcaaga acagggtcca gaacatggct ctttatgccg
      961 acgttagtgg aaaacaattt cctgtaaccc gcggccagga tgtgggtcga tatcaggtgt
     1021 cctggagcct ggagcacaag agcgctcatg cgggcaccta tgaggtcaga ttctttgatg
     1081 aagagtccta tagcctccta aggaaggctc agagaaataa tgaggacgtt tctatcattc
     1141 cacccctgtt cacagtcagt gtggaccatc ggggtacctg gaatgggcct tgggtctcca
     1201 cagaggtgct ggctgcagta attggcatag tgatctacta cctagctttc agtgcaaaga
     1261 gccacatcca ggccggatcc gactacaaag accatgacgg tgattataaa gatcatgaca
     1321 tcgactacaa ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct
     1381 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     1441 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     1501 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     1561 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     1621 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     1681 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     1741 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     1801 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     1861 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     1921 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact
     1981 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     2041 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     2101 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     2161 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     2221 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     2281 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2341 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2401 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     2461 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2521 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2581 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2641 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2701 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2761 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2821 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2881 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2941 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3001 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3061 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3121 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3181 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3241 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3301 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3361 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3421 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3481 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3541 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3601 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3661 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3721 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3781 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3841 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     3901 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     3961 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4021 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4081 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4141 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4201 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4261 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4321 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4381 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4441 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4501 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4561 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4621 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4681 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4741 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4801 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     4861 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4921 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4981 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5041 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     5101 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5161 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5221 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5281 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5341 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5401 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5461 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5521 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5581 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5641 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5701 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5761 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5821 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5881 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5941 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6001 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6061 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6121 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6181 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6241 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6301 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6361 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     6421 agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     6481 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     6541 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
