LOCUS       V046170                 9642 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046170
VERSION     V046170
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..4322
                     /label="Anks1(NM_001286040)"
                     /note="Anks1(NM_001286040)"
                     /gene="Anks1"
     CDS             4329..4394
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4418..5006
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5072..5120
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5322..5750
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5764..6093
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6160..6954
                     /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    7130..7251
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7300..7316)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7324..7340
                     /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(7348..7378)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7393..7414
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7702..8290)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8461..9321)
                     /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(9322..9426)
                     /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 ccaccatggg gaaggagcag gagctgctgg
      781 aggcggcccg caccgggcac ctcccggccg tggagaagct gctgtccggg aagcggctct
      841 cctccggctt cggcggcggc ggcggcggct caggcagcgg cggcggctcc gggggcggcg
      901 gcctcggttc ttctagccac ccgctctcca gtctgctgag catgtggaga gggccgaacg
      961 tgaactgtgt agatagcacc ggctacacgc cactacacca cgctgctttg aatggacaca
     1021 gggatgtggt cgaggtcctt ctgaggaacg acgcgctgac caacgttgct gactcgaaag
     1081 gctgctaccc gctgcacttg gcagcgtgga aaggagacgc ccagatagta cgtctgctca
     1141 tccagcaagg gccctcacac accagagtca acgaacagaa tgctcttgag atcagagaac
     1201 tcaaaaagta cggccccttt gacccttata tcaatgccaa gaacaacgac aatgagacgg
     1261 ccctgcactg cgcggcccag tacggccaca cggaggtcgt gaaggctctg ctggaggagc
     1321 tcacggaccc caccatgcgc aacaacaagt ttgaaacgcc cctggacttg gcggcgctgt
     1381 acgggcggct ggaggtggtg aagctgctgc tgggtgcaca ccccaatctc ctgagctgca
     1441 gcactcggaa acacacgccc ctgcacttgg cagcaaggaa tggccacaag gccgtggtgc
     1501 aggtcctgct ggacgccggc atggatagca actatcagac ggagatgggc agtgctttgc
     1561 atgaggctgc tttgtttggc aagaccgatg tggtgcaaat cctgctggct gcaggaatcg
     1621 atgttaacat aaaggataac cgtggcctga ctgcccttga cactgtccgg gatctgccct
     1681 ctcagaagag ccagcagata gccgccctga ttgaagatca catgaccgga aaaagaagcg
     1741 tgaaagaagt ggacaggacc tccacagccc agctacctct cctctccaac acagacgcca
     1801 tagcaccaat gtctcagggg agtatggaga agacagtgac tgagctgatc cttcattttg
     1861 acacacatgc tgacgaggag ggcccctacg aggcactgta caacgctgtg tcctgccatt
     1921 ccttggacag cacggccagt ggacgatcat cagacagaga ctccatgaac aaggaggccg
     1981 aggccaccgg aacaagggct gctggagtga ggcctaggga acgtccgccg cctccagcaa
     2041 aaccgccacc cgatgaagag gaggaagaac gcgtagataa gaagtatttt cccttggcag
     2101 cctctgaggg cctggctgtg agacccagga ttcagagtag tgctccccag gaagaggagg
     2161 agcacccata tgagctgctg ctcacggcag aaaccaagaa gctggggacc acagacggga
     2221 ggactgaaga ccacaggcag agcggcagcg gccggagcca ggactctgtg gaggggcagg
     2281 acgggcaggt cccagagcag ttctcaggtc tcctccacgg ctcctcccca gtgtgtgagg
     2341 tggggcagga ccctttccag ctgctcacgg cccccagcca gagccacccg gagtcctccc
     2401 agcaggacgc ctgccacgag gccagcatgc agctagagga gccaggcgtg caaggcactg
     2461 aacccccgca gcccggcgtc cctgaccaaa gcaagagagt gggcttgccg gcaggcctga
     2521 cagccctggc cagcagaaca tacctcgatg cattgactca cacagtacct ctgcgccctg
     2581 ctggtgccga ggaagaagac cagagcgggc ccagaagcag agcccctccc accagcaagc
     2641 ccaaagctga gctcaaactc agccgcagtt tgtctaagtc tgactctgac ctcctgacct
     2701 gctcccccac ggaggacgcc acaatgggga gtcgcagcga gtccttgtcc aactgcagca
     2761 tcgggaagaa gaggttggag aaatcacctt cctttgcctc agagtgggat gagattgaga
     2821 aaatcatgag ctccattggg gaagggattg acttctctca ggagcagcag aagatctcag
     2881 gttctcggac gctggagcag agcgtcgggg agtggctgga gtcgattggc ctgcagcagt
     2941 acgagagcaa gctgctgctg aacggctttg atgatgtccg cttcctgggg tctaatgtga
     3001 tggaagagca ggacctgagg gagattggca tcagtgaccc acagcaccgg cggaagctgc
     3061 tgcaggctgc acgctccttg cccaaggtga aggcccttgg ctatgacggg gtcagcccca
     3121 cctcagtgcc ctcatggctg gactcgctgg ggctgcagga ctatgtccac tctttcctgt
     3181 ccagtggcta cagctccatc gacactgtga agaacctctg ggagctagag cttgtcaatg
     3241 tcctgaaagt tcacctgctg ggccaccgca agcgcattat cgcctctcta gcagacaggc
     3301 cctatgagga gccaccccag aagcctccaa ggttttccca gctaagatgc caggatttaa
     3361 tctcccagac atcgtccccg ctgagccaga atgattcctg cacagggcgg tcggcagacc
     3421 tgctgctgcc ctcagcagac acaagcagga ggcggcacga cagcctgccc gaccctggga
     3481 ccgcctcccg agcagaccgc ttccgagtcc aggaggagcc cagtgagacc aagctgaccc
     3541 tccgtccacc cagcctagcc gcgccctatg ctcctgttca gagctggcag caccagccag
     3601 aaaagctcat ctttgagtcc tgcggttatg aggccaatta tctgggttcc atgctgatca
     3661 aagatctgcg agggacagag tccacccaag atgcctgtgc caaaatgcgg aagtcaacgg
     3721 agcacatgaa gaagatcccc accatcatcc tgtccatcac gtacaaaggt gtcaagttca
     3781 tcgatgcttc caacaagaac gtcattgcgg agcacgagat ccggaacatt tcctgcgccg
     3841 cgcaggaccc agaggacctg tgtaccttcg cgtacatcac caaggacctg cagacgagcc
     3901 accactactg ccacgttttc agcacagtgg atgtgaacct gacctacgag atcatcctga
     3961 ccctggggca ggcatttgag gtggcctatc agctggccct ccaagcccag aaatccagga
     4021 ccatggcggc ctctgcagct tcgatgattg agacaaaatc ctccaaacca gtgcctaagc
     4081 ctcgggtcgg catgaggaag tctgcagtgc cggtgccccc cgactctcgc tgttgtcact
     4141 gtcacacctg caccactcac cgtccttcct acctaccgct gccgtccgtt agtcctggag
     4201 tcaagctgga accacctgat tctgaccaag aggccccgtc ccacgctagt gtttcctgga
     4261 tcgtggaccc aaagccagac tcgaagcgga gcctcagcac caagtacgag accactatct
     4321 tcggatccga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     4381 atgacgatga caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat
     4441 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     4501 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     4561 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     4621 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     4681 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     4741 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     4801 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     4861 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     4921 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     4981 gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag
     5041 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     5101 gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     5161 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     5221 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     5281 catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     5341 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     5401 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5461 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     5521 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5581 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5641 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     5701 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     5761 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5821 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     5881 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5941 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     6001 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     6061 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     6121 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     6181 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     6241 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     6301 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     6361 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     6421 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     6481 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     6541 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     6601 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     6661 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     6721 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     6781 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     6841 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     6901 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     6961 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     7021 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     7081 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     7141 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     7201 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     7261 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     7321 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     7381 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     7441 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     7501 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     7561 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     7621 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7681 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7741 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7801 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7861 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     7921 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7981 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     8041 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     8101 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     8161 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     8221 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     8281 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     8341 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     8401 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     8461 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     8521 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     8581 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     8641 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     8701 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     8761 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     8821 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     8881 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8941 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     9001 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     9061 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     9121 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     9181 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     9241 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     9301 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     9361 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     9421 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     9481 cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     9541 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     9601 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg
//
