LOCUS       V034602                 7194 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034602
VERSION     V034602
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..1874
                     /label="MAF(NM_001031804)"
                     /note="MAF(NM_001031804)"
                     /gene="MAF"
     CDS             1881..1946
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1970..2558
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2624..2672
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2874..3302
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3316..3645
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3712..4506
                     /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    4682..4803
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4852..4868)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4876..4892
                     /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(4900..4930)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4945..4966
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5254..5842)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6013..6873)
                     /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(6874..6978)
                     /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 atcagaactg gcaatgagca
      781 actccgacct gcccaccagt cccctggcca tggaatatgt taatgacttc gatctgatga
      841 agtttgaagt gaaaaaggaa ccggtggaga ccgaccgcat catcagccag tgcggccgtc
      901 tcatcgccgg gggctcgctg tcctccaccc ccatgagcac gccgtgcagc tcggtgcccc
      961 cttcccccag cttctcggcg cccagcccgg gctcgggcag cgagcagaag gcgcacctgg
     1021 aagactacta ctggatgacc ggctacccgc agcagctgaa ccccgaggcg ctgggcttca
     1081 gccccgagga cgcggtcgag gcgctcatca gcaacagcca ccagctccag ggcggcttcg
     1141 atggctacgc gcgcggggcg cagcagctgg ccgcggcggc cggggccggt gccggcgcct
     1201 ccttgggcgg cagcggcgag gagatgggcc ccgccgccgc cgtggtgtcc gccgtgatcg
     1261 ccgcggccgc cgcgcagagc ggcgcgggcc cgcactacca ccaccaccac caccacgccg
     1321 ccggccacca ccaccacccg acggccggcg cgcccggcgc cgcgggcagc gcggccgcct
     1381 cggccggtgg cgctgggggc gcgggcggcg gtggcccggc cagcgctggg ggcggcggcg
     1441 gcggcggcgg cggcggaggc ggcgggggcg cggcgggggc ggggggcgcc ctgcacccgc
     1501 accacgccgc cggcggcctg cacttcgacg accgcttctc cgacgagcag ctggtgacca
     1561 tgtctgtgcg cgagctgaac cggcagctgc gcggggtcag caaggaggag gtgatccggc
     1621 tgaagcagaa gaggcggacc ctgaaaaacc gcggctatgc ccagtcctgc cgcttcaaga
     1681 gggtgcagca gagacacgtc ctggagtcgg agaagaacca gctgctgcag caagtcgacc
     1741 acctcaagca ggagatctcc aggctggtgc gcgagaggga cgcgtacaag gagaaatacg
     1801 agaagttggt gagcagcggc ttccgagaaa acggctcgag cagcgacaac ccgtcctctc
     1861 ccgagttttt catgggatcc gactacaaag accatgacgg tgattataaa gatcatgaca
     1921 tcgactacaa ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct
     1981 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2041 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2101 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2161 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2221 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2281 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2341 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     2401 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     2461 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     2521 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact
     2581 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     2641 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     2701 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     2761 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     2821 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     2881 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2941 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3001 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     3061 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3121 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3181 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3241 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3301 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3361 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3421 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3481 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3541 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3601 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3661 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3721 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3781 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3841 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3901 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3961 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4021 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4081 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4141 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4201 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4261 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4321 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4381 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4441 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4501 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     4561 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4621 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4681 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4741 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4801 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4861 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4921 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4981 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5041 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5101 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5161 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5221 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5281 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5341 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5401 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     5461 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5521 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5581 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5641 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     5701 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5761 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5821 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5881 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5941 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6001 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6061 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6121 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6181 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6241 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6301 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6361 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6421 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6481 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6541 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6601 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6661 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6721 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6781 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6841 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6901 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6961 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     7021 agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     7081 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     7141 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
