LOCUS       V038734                 7380 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038734
VERSION     V038734
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..2060
                     /label="AP2M1(NM_004068)"
                     /note="AP2M1(NM_004068)"
                     /gene="AP2M1"
     CDS             2067..2132
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2156..2744
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2810..2858
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3060..3488
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3502..3831
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3898..4692
                     /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    4868..4989
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5038..5054)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5062..5078
                     /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(5086..5116)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5131..5152
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5440..6028)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6199..7059)
                     /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(7060..7164)
                     /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 ccaccatgat tggaggctta ttcatctata
      781 atcacaaggg ggaggtgctc atctcccgag tctaccgaga tgacatcggg aggaacgcag
      841 tggatgcctt tcgggtcaat gttatccatg cccggcagca ggtgcgcagc cccgtcacca
      901 acattgctcg caccagcttc ttccacgtta agcggtccaa catttggctg gcagcagtca
      961 ccaagcagaa tgtcaacgct gccatggtct tcgaattcct ctataagatg tgtgacgtga
     1021 tggctgccta ctttggcaag atcagcgagg aaaacatcaa gaacaatttt gtgctcatat
     1081 atgagctgct ggatgagatt ctagactttg gctacccaca gaattccgag acaggcgcgc
     1141 tgaaaacctt catcacgcag cagggcatca agagtcagca tcagacaaaa gaagagcagt
     1201 cacagatcac cagccaggta actgggcaga ttggctggcg gcgagagggt atcaagtatc
     1261 gtcggaatga gctcttcctg gatgtgctgg agagtgtgaa cctgctcatg tccccacaag
     1321 ggcaggtgct gagtgcccat gtgtcgggcc gggtggtgat gaagagctac ctgagtggca
     1381 tgcctgaatg caagtttggg atgaatgaca agattgttat tgaaaagcag ggcaaaggca
     1441 cagctgatga aacaagcaag agcgggaagc aatcaattgc cattgatgac tgcaccttcc
     1501 accagtgtgt gcgactcagc aagtttgact ctgaacgcag catcagcttt atcccgccag
     1561 atggagagtt tgagcttatg aggtatcgca caaccaagga catcatcctt cccttccggg
     1621 tgatcccgct agtgcgagaa gtgggacgca ccaaactgga ggtcaaggtg gtcatcaagt
     1681 ccaactttaa accctcactg ctggctcaga agatcgaggt gaggatccca accccactga
     1741 acacaagcgg ggtgcaggtg atctgcatga aggggaaggc caagtacaag gccagcgaga
     1801 atgccatcgt gtggaagatc aagcgcatgg caggcatgaa ggaatcgcag atcagcgcag
     1861 agattgagct tctgcctacc aacgacaaga agaaatgggc tcgacccccc atttccatga
     1921 actttgaggt gccattcgcg ccctctggcc tcaaggtgcg ctacttgaag gtgtttgaac
     1981 cgaagctgaa ctacagcgac catgatgtca tcaaatgggt gcgctacatt ggccgcagtg
     2041 gcatttatga aactcgctgc ggatccgact acaaagacca tgacggtgat tataaagatc
     2101 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca
     2161 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2221 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2281 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     2341 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     2401 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     2461 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     2521 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     2581 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     2641 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     2701 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     2761 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     2821 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     2881 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     2941 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     3001 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     3061 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3121 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3181 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     3241 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3301 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3361 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3421 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3481 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3541 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3601 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3661 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3721 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3781 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3841 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     3901 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3961 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4021 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4081 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4141 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4201 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4261 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4321 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4381 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4441 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4501 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4561 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4621 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4681 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     4741 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4801 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4861 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4921 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4981 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5041 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5101 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5161 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5221 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5281 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5341 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5401 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5461 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5521 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5581 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     5641 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5701 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5761 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5821 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5881 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5941 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6001 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6061 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6121 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6181 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6241 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6301 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6361 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6421 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6481 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6541 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6601 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6661 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6721 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6781 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6841 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6901 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6961 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7021 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7081 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7141 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     7201 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     7261 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     7321 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
