LOCUS       V031916                 9208 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031916
VERSION     V031916
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"
     CDS             706..3864
                     /label="Ankrd28(NM_001024604)"
                     /note="Ankrd28(NM_001024604)"
                     /gene="Ankrd28"
     CDS             3871..3960
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3984..4572
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4638..4686
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4888..5316
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5330..5659
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5726..6520
                     /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    6696..6817
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6866..6882)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6890..6906
                     /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(6914..6944)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6959..6980
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7268..7856)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8027..8887)
                     /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(8888..8992)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggc gttcctcaaa
      721 ctccgcgacc agccatcact ggtgcaagct atatttaatg gtgatccaga tgaagtccga
      781 gcactaatat ttaagaaaga agatgttaac tttcaggaca atgagaagag aacccctttg
      841 catgctgctg cctatcttgg tgacgcagaa atcattgagc tgcttatttt atctggagct
      901 agagttaatg ccaaagacag caaatggttg acacctttac acagagcagt tgcatcgtgt
      961 agtgaggaag cagttcagat acttttgaag cattctgcag atgttaatgc tcgagataaa
     1021 aattggcaga cccctttaca catagctgct gctaataaag ctgtaaagtg cgctgaatct
     1081 ttggtacctc ttctgagcaa tgtgaacgtg tccgaccgag caggaagaac tgcattacat
     1141 catgcagctt tcagtgggca tggtgagatg gtcaaactac tcttgtccag aggtgccaat
     1201 ataaatgctt ttgacaagaa agataggcgt gcaatccatt gggcagcata tatgggtcac
     1261 attgaagtag taaaattact tgtatctcat ggagctgaag tgacatgtaa ggataagaag
     1321 tcttacacac ctcttcatgc agcagcctct agtggaatga tcagtgtagt caaatatctt
     1381 ctagatcttg gagttgatat gaatgaacca aatgcctatg gaaacacacc tcttcatgtg
     1441 gcctgttaca atggacaaga tgttgttgtg aatgagctca tagactgtgg tgctaatgtc
     1501 aatcagaaga atgagaaagg gttcactcct ttgcactttg ctgctgcatc aacacatgga
     1561 gcactgtgtt tagaacttct ggttggcaat ggagctgatg tcaatatgaa gagtaaagat
     1621 gggaaaaccc cactacatat gactgccctc cacggtagat tctccagatc acaaacgatt
     1681 atccagagtg gagctgtaat agactgtgaa gataagaatg ggaacacccc tttgcatata
     1741 gcagcccggt atggccatga gcttctaatc aacactctta ttacaagtgg tgctgacact
     1801 gcaaagcgtg gtatacatgg aatgttccca ctccatttgg cagccttaag tggtttttca
     1861 gattgctgca ggaaactcct ttcttcagga tttgacatag ataccccaga tgattttggc
     1921 agaacctgtc tacacgcagc tgcagctgga ggaaatttgg agtgcctaaa ccttctgctg
     1981 aatactggtg cagacttcaa caaaaaggat aaatttggaa gatctccact gcactatgct
     2041 gctgccaact gcaattacca gtgcctgttt gctcttgtgg gatcaggagc aagtgtgaat
     2101 gaccttgatg agagaggctg cacacccctg cactatgcag ctacatctga cacagacggc
     2161 aagtgcctgg aatacttact aagaaatgat gccaacccag ggatccggga caaacaaggc
     2221 tataatgcag ttcactactc agctgcgtat ggtcaccgtc tgtgtcttca actgattgca
     2281 agtgaaactc ctttagatgt tttaatggaa acctcgggaa cagacatgtt gagtgattca
     2341 gataatagag caacaataag ccctttacat ttggctgctt atcatggtca ccatcaagcc
     2401 ctggaagtgt tggtacagtc tttgttagat cttgatgtaa gaaatagtag tggtagaaca
     2461 cccttagatc ttgcagcctt taagggccat gttgaatgtg tggatgtact cattaatcag
     2521 ggagcttcaa tcctcgtaaa agattacgtt ttgaagagaa cacctatcca tgcagcagca
     2581 acaaatggtc attcagaatg cttacggtta ttaataggaa atgcagagcc acagaatgca
     2641 gtagatattc aagatggaaa tggacagaca cctcttatgc tgtcggttct caatgggcac
     2701 acggactgtg tatactcact actgaacaaa ggagcaaatg tagatgccaa agacaagtgg
     2761 ggaaggacag cgttgcacag aggggcagtg acaggccatg aagagtgtgt agacgcatta
     2821 cttcaacatg gtgctaagtg cttactacgg gacagccggg gtcggacacc catacacttg
     2881 tcagcagcct gtggacacat tggagttctt ggagcccttt tgcagtcagc aacatctgtg
     2941 gatgcaaatc cagccgtagt agacaatcat gggtatacag cacttcactg ggcttgctac
     3001 aatggtcatg agacatgtgt ggagctactt ttagaacagg atgttttcca gaaaatagat
     3061 ggaaatgctt tcagccccct gcattgtgcc gtgatcaacg acaatgaagg tgcagctgag
     3121 atgctgattg attccttagg agccagcatt gtgaatgcca cagactcaaa gggaagaact
     3181 cctctccatg ccgctgcctt cacagaccat gtggagtgtt tacaactgct gctcagccag
     3241 aatgctcagg tcaattccgc agactctacc ggaaaaacac ctctcatgat ggctgcagaa
     3301 aacggacaaa ccaatacagt tgagatgctg gttagcagtg ctagtgcaga tctgacatta
     3361 caagataaga gtaaaaacac cgcccttcat ttggcttgtg gcaagggtca tgaaactagt
     3421 gccttgttaa ttctggaaaa gataacagat agaaacctca tcaatgcaac caacgcagcc
     3481 ttgcaaacac ctctgcatgt tgctgcccga aatgggctca cgatggttgt tcaagagctc
     3541 ctggggaaag gagccagtgt gcttgctgtt gatgagaatg gctacacccc agctttggcc
     3601 tgtgctccca ataaggatgt ggctgactgc ctggctctca ttttggccac catgatgcct
     3661 gtctcgtcaa gtagcccatt gacatccctg acattcaatg ccattaaccg ttataccaac
     3721 acctcaaaaa cagttagctt tgaagccttg cctatcatga ggaatgaagc cagctcctac
     3781 tgcagtttta acaacattgg tggcgagcag gagtacttgt acaccgacgt ggatgagctc
     3841 aacgactctg actccgagac ctacctcgag gaacaaaaac taatatcaga ggaagatttg
     3901 gaacagaaac taatcagtga agaagattta gagcaaaagt taatttccga ggaggactta
     3961 tagtgagggc ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga
     4021 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     4081 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     4141 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     4201 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     4261 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     4321 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     4381 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     4441 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     4501 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     4561 ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga
     4621 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt
     4681 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     4741 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     4801 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc
     4861 gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     4921 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4981 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     5041 ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     5101 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5161 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     5221 ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa
     5281 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     5341 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5401 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5461 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     5521 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5581 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     5641 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     5701 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     5761 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     5821 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     5881 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     5941 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     6001 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     6061 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     6121 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     6181 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     6241 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     6301 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     6361 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     6421 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     6481 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg
     6541 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     6601 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     6661 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     6721 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6781 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     6841 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6901 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6961 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     7021 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     7081 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     7141 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7201 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7261 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7321 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7381 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7441 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
     7501 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7561 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7621 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7681 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     7741 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7801 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7861 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7921 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7981 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     8041 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     8101 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     8161 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     8221 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     8281 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8341 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8401 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8461 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8521 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8581 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8641 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8701 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8761 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8821 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8881 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8941 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     9001 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac
     9061 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     9121 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     9181 cgacaattgc atgaagaatc tgcttagg
//
