LOCUS       V021246                10082 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021246
VERSION     V021246
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             683..4705
                     /label="ACIN1(NM_014977)"
                     /note="ACIN1(NM_014977)"
                     /gene="ACIN1"
     CDS             4745..4834
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    4858..5446
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5512..5560
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5762..6190
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6204..6533
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6600..7394
                     /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    7570..7691
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7740..7756)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7764..7780
                     /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(7788..7818)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7833..7854
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8142..8730)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8901..9761)
                     /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(9762..9866)
                     /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 ggatccgcca ccatgtggag acggaaacat ccgaggacat ccggaggaac
      721 ccggggagtt ctgagtggta atcgaggggt agagtatggc agtgggcggg gtcatctcgg
      781 tacgttcgaa gggcgttggc ggaaattacc gaagatgccc gaagccgtcg ggacggaccc
      841 gagtacctca cgcaagatgg cggagctgga ggaggtgact ctggacggga agcctcttca
      901 ggcgctgcgg gtgaccgacc tgaaggccgc actggagcag cgaggcctag ccaagagcgg
      961 gcagaagagt gccctggtca agcggctcaa aggggctcta atgctagaaa atttacagaa
     1021 acactcaaca ccccatgctg cattccagcc aaattcccag attggtgagg aaatgagcca
     1081 gaacagtttc ataaaacagt atctggaaaa gcagcaggag ctacttaggc agcgtctgga
     1141 acgtgaagct cgagaagctg cagaacttga agaagcttca gctgagtcgg aggacgagat
     1201 gatccatcct gagggagtgg cttccctgct gcctcctgac tttcagagca gcctggagag
     1261 accagagctg gagctcagca gacattcgcc cagaaaaagc tcctcaattt ctgaagagaa
     1321 aggtgactct gatgatgaga aaccaaggaa aggagaaaga cgatcatcta gggtcagaca
     1381 ggcaagagca gctaaactgt ctgagggcag ccaacctgct gaggaggaag aggatcaaga
     1441 aacaccttcc agaaacctaa gggtcagagc agatcgaaat ttgaaaacag aggaggaaga
     1501 agaggaggag gaggaggagg aagaagatga tgaagaagag gaaggtgatg atgagggaca
     1561 aaaatctagg gaggcaccaa tcctgaaaga gtttaaggaa gaaggggaag agatacctag
     1621 agtaaaacca gaggagatga tggatgagag acccaaaaca agatcccagg aacaggaggt
     1681 gttagagaga ggagggagat ttacaagatc ccaggaagag gctagaaaaa gtcatctggc
     1741 cagacagcag caggagaagg aaatgaaaac aacatctccc cttgaggagg aagaaagaga
     1801 aataaaatct tcacaaggct taaaggaaaa atcgaagtct ccttcccctc ctcgactgac
     1861 tgaagatcga aagaaggcct cacttgtagc gctgccagag caaactgcca gcgaggagga
     1921 gactcctcca cctttactaa caaaggaagc atcttctcca ccacctcatc cacagctcca
     1981 tagcgaagaa gaaatagagc ccatggaagg cccagccccc cctgtcctca ttcagttatc
     2041 tcctcctaat acagatgctg acaccaggga gctattagta tctcagcata ctgtccagtt
     2101 ggtaggaggc ctgtctcctt tgtcaagtcc ttcagacacc aaagcagaat ctccagcaga
     2161 gaaagtgcca gaggagagtg tcctgcctct ggttcagaaa agcacactgg ctgactactc
     2221 agcccagaag gatcttgaac ctgagtcaga cagatctgct cagcccctcc ctctaaaaat
     2281 tgaggaatta gcactggcca aaggaatcac tgaagaatgt ctgaaacagc catctttgga
     2341 acagaaggaa ggcagaagag cttctcatac ccttctccca agccacagat tgaaacagtc
     2401 agctgattca tcctctagcc ggtcctcctc atcttcctcc tccagttcta gatcaagatc
     2461 tcgctctcct gacagttcag gttctcggtc tcattcaccg ctcagatcca agcagagaga
     2521 tgtagcccag gcacgtactc atgccaaccc tcgtggtaga cccaagatgg gctccagatc
     2581 aacatcagag tccagatcaa ggtcacgttc acgttctcgt tcagcatcaa gcaacagcag
     2641 aaaatctctg agccctggag tctccaggga cagcagcacc agctatactg aaaccaaaga
     2701 tccctcttct ggtcaggagg ttgcaactcc accagtgcca caactgcagg tctgtgagcc
     2761 aaaggagagg acttccacct cctcatcctc tgtccaagca aggcgtctga gtcagcctga
     2821 atcagctgaa aagcatgtga cccagaggtt acagcctgag cgggggagcc caaagaagtg
     2881 tgaagctgaa gaggcagagc caccagctgc cacacagccc caaacctcag agactcagac
     2941 ctctcatctg ccagaatcag aaagaattca tcacactgtt gaggagaagg aggaagtgac
     3001 catggacaca agtgaaaaca gacctgaaaa tgatgttcca gaacctccca tgcctattgc
     3061 agaccaagtc agcaatgatg accgcccgga gggcagtgtt gaagatgagg agaagaaaga
     3121 gagctcgctg cccaaatcat tcaagaggaa gatctccgtt gtctcagcta ccaagggggt
     3181 gccagctgga aacagtgaca cagagggggg ccagcctggt cggaaacgac gctggggagc
     3241 cagcacagcc accacacaga agaaaccttc catcagtatc accactgaat cactaaagag
     3301 cctcatcccc gacatcaaac ccctggcggg gcaggaggct gttgtggatc ttcatgctga
     3361 tgactctcgc atctctgagg atgagacaga gcgtaatggc gatgatggga cccatgacaa
     3421 ggggctgaaa atatgccgga cagtcactca ggtagtacct gcagagggcc aggagaatgg
     3481 gcagagggaa gaagaggaag aagagaagga acctgaagca gaacctcctg tacctcccca
     3541 ggtgtcagta gaggtggcct tgcccccacc tgcagagcat gaagtaaaga aagtgacttt
     3601 aggagatacc ttaactcgac gttccattag ccagcagaag tccggagttt ccattaccat
     3661 tgatgaccca gtccgaactg cccaggtgcc ctccccaccc cggggcaaga ttagcaacat
     3721 tgtccatatc tccaatttgg tccgtccttt cactttaggc cagctaaagg agttgttggg
     3781 gcgcacagga accttggtgg aagaggcctt ctggattgac aagatcaaat ctcattgctt
     3841 tgtaacgtac tcaacagtag aggaagctgt tgccacccgc acagctctgc acggggtcaa
     3901 atggccccag tccaatccca aattcctttg tgctgactat gccgagcaag atgagctgga
     3961 ttatcaccga ggcctcttgg tggaccgtcc ctctgaaact aagacagagg agcagggaat
     4021 accacggccc ctgcaccccc cacccccacc cccggtccag ccaccacagc acccccgggc
     4081 agagcagcgg gagcaggaac gggcagtgcg ggaacagtgg gcagaacggg aacgggaaat
     4141 ggagcggcgg gagcggactc gatcagagcg tgaatgggat cgggacaaag ttcgagaagg
     4201 gccccgttcc cgatcaaggt cccgtgaccg ccgccgcaag gaacgtgcga agtctaaaga
     4261 aaagaagagt gagaagaaag agaaagccca ggaggaacca cctgccaagc tgctggatga
     4321 ccttttccga aagaccaagg cagctccctg catctattgg ctcccactga ctgacagcca
     4381 gatcgttcag aaagaggcag agcgggccga acgggccaag gagcgggaga agcggcgaaa
     4441 ggagcaagaa gaagaagagc aaaaggagcg ggagaaggaa gccgagcggg aacggaaccg
     4501 acagctggag cgagagaaac gtcgggagca cagtcgggag agggacaggg agagagagag
     4561 agaaagggag cgggacaggg gggaccgaga tcgggatagg gaaagggacc gagaacgagg
     4621 cagggaaagg gatcgcaggg acaccaagcg ccacagcaga agccggagtc ggagcacacc
     4681 tgtgcgggac cggggtgggc gccgcgaatt ctgcagatat ccagcacagt ggcggccgct
     4741 cgaggaacaa aaactaatat cagaggaaga tttggaacag aaactaatca gtgaagaaga
     4801 tttagagcaa aagttaattt ccgaggagga cttatagtga gggcccgata tctcgacaat
     4861 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     4921 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     4981 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     5041 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     5101 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     5161 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     5221 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     5281 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     5341 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     5401 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga
     5461 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     5521 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     5581 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     5641 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     5701 gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc
     5761 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     5821 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     5881 acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt
     5941 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     6001 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     6061 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     6121 taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     6181 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     6241 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     6301 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     6361 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     6421 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     6481 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     6541 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     6601 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     6661 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     6721 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     6781 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     6841 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     6901 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     6961 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     7021 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     7081 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     7141 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     7201 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     7261 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     7321 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     7381 acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac
     7441 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     7501 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     7561 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     7621 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     7681 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     7741 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     7801 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     7861 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     7921 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     7981 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     8041 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     8101 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     8161 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     8221 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     8281 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     8341 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     8401 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     8461 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     8521 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     8581 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     8641 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     8701 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     8761 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     8821 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     8881 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     8941 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     9001 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     9061 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     9121 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     9181 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     9241 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     9301 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     9361 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     9421 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     9481 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     9541 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     9601 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     9661 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     9721 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     9781 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     9841 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     9901 ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg
     9961 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
    10021 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
    10081 gg
//
