LOCUS       V026524                 8622 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026524
VERSION     V026524
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..3302
                     /label="Akap4(NM_009651)"
                     /note="Akap4(NM_009651)"
                     /gene="Akap4"
     CDS             3309..3374
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3398..3986
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4052..4100
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4302..4730
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4744..5073
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5140..5934
                     /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    6110..6231
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6280..6296)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6304..6320
                     /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(6328..6358)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6373..6394
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6682..7270)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7441..8301)
                     /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(8302..8406)
                     /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 tgcctactgt ggtactacaa
      781 cgatgtctga cgatattgac tggttacaca gccgcagagg cgtgtgcaag gtagatcttt
      841 acagtccaaa aggacaacaa gatcaggacc gaaaagtgat atgctttgtg gatgtgtcta
      901 ccttgaatgt ggaagataaa gattccaagg gtgctgctgg ttccaggtca gaaggcgagt
      961 taaatctgga gactctggaa gaaaaagaga ttatcgtaat caaggacact gaaaaacaag
     1021 accagtctaa gacggaggga tctgtgtgcc ttttcaaaca agctccctct gatcctataa
     1081 gtgtcctcaa ctggctcctc aatgatctcc agaagtatgc cttgggtttc caacatgcat
     1141 taagcccctc agcctctagc tgtaaacata aagtgggaga cctagagggt gactatagca
     1201 aaataccatc tgagaactgc tacagtgtgt acgctgatca agtaaacttt gattatttga
     1261 acaaaggacc tcaaaacctt cgcctagaaa tggcggcatc caaaaacacc aacaataacc
     1321 aaagtccctc aaatcctgca accaaatctc ctagcaatca gaggtcagtt gccactcctg
     1381 agggagaatg ttctatggac gacctttcct tctatgtcaa ccgattgtca tctctggtaa
     1441 tccaaatggc ccgtaaggaa atcaaggaca aattggaagg tggaagcaaa tgtctccatc
     1501 attccatgta tacatcagga gataaaggga aaaccagccc ccggagtgct gtcagcaaaa
     1561 ttgcttcgga gatggcccat gaagctgttg aactgacctc atcagaaatg cgtggcaatg
     1621 gagaggattg cagggatggc cggaaaacct ttttgtatag tgaaatgtgt aacaagaaca
     1681 agtgtggcga aaagcagcag atgtgcccaa aagacagcaa agaatttgcg gattccatca
     1741 gcaaggggct tatggtttat gcaaatcaag tagcatctga catgatggtc tctgttatga
     1801 aaactttgaa agtacacagc tgtgggaagc caattccggc ttgtgtggtc ctgaagaggg
     1861 tactattgaa gcacaccaag gaaattgtat ctgatctgat tgattcatgt atgaaaaact
     1921 tgcataacat aacaggagtc ctgatgacag actcagactt cgtttctgct gtcaagagga
     1981 atcttttcaa tcatggtaaa caaaatgcag cggacatcat ggaggccatg ctaaagcgtc
     2041 tggtcagtgc tcttcttggt gagaagaagg agactaagtc tcaaagtctg gcctatgcaa
     2101 ccttaaaagc tggaactaat gatccgaaat gcaagaatca gagccttgag ttctcagcta
     2161 tgaaagctga aatgaaagga aaagataaat gcaaatccaa agcagatcca tgctgcaaat
     2221 cgttgacaag tgctgagaga gtcagcgagc atatcctcaa agagagcctt actatgtgga
     2281 acaaccagaa gcaaggaaac caaggcaaag tgactaacaa agtttgctgc accagtaaag
     2341 atgagaagag agaaaagatc agtccttcca cagattcact ggccaaggat ctaattgtct
     2401 ctgcccttat gctcattcaa taccatctga cccaacaagc caagggcaaa gatccatgtg
     2461 aagaggagtg ccctggttcc tccatggggt atatgtccca aagtgcacaa tacgaaaaat
     2521 gtggaggtgg ccaaagttct aaatcacttt caatgaagca ttttgaaact cgtggagctc
     2581 ctggaccatc tacatgtatg aaggaaaatc aactggagtc ccagaagatg gatatgtcaa
     2641 acatggttct gtccctgatt cagaaactcc tgagtgagag ccctttcagt tgcgatgaac
     2701 taactgaaag tgacaataag cgttgttgtg atcctagatc aagcaaagca gctcccatgg
     2761 ccaagagacc tgaagagcaa tgccaagaca atgcagaact agacttcatc agtgggatga
     2821 agcaaatgaa ccgccagttt atagatcagc tggtagaatc cgtgatgaaa ctctgcctga
     2881 tcatggctaa gtacagcaac aatggagcag ccctggctga gctggaagaa caagcagccc
     2941 tggtaggcag tggctccaga tgtggccgtg atgctatgat gtcacaaaat tattctgaaa
     3001 ctcctggccc tgaagttatt gtcaacaatc agtgctctac aactaacttg cagaagcagc
     3061 tgcaggctgt cctgcaatgg attgcagcct ctcaattcaa tgtgcccatg ctctacttca
     3121 tgggagacga tgatggacaa ctagagaagc tccctgaagt ttcagctaag gctgcagaga
     3181 aggggtacag tgtaggagat cttcttcagg aggtcatgaa gtttgccaaa gaacgacaac
     3241 tggatgaagc cgtgggcaac atggctagaa agcagctgct agactggctt ctcgctaacc
     3301 tgggatccga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     3361 atgacgatga caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat
     3421 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     3481 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     3541 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     3601 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3661 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3721 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3781 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     3841 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3901 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3961 gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag
     4021 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     4081 gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     4141 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     4201 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     4261 catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     4321 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4381 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4441 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     4501 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4561 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4621 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     4681 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4741 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4801 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4861 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4921 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4981 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     5041 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     5101 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     5161 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     5221 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     5281 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     5341 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     5401 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     5461 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5521 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5581 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5641 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     5701 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5761 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5821 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5881 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5941 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     6001 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     6061 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     6121 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6181 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     6241 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6301 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6361 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6421 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6481 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6541 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6601 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6661 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6721 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6781 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6841 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     6901 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6961 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7021 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7081 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     7141 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7201 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7261 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7321 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7381 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7441 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7501 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7561 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7621 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7681 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7741 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7801 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7861 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7921 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7981 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8041 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8101 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8161 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8221 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8281 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8341 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8401 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     8461 cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     8521 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     8581 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg
//
