LOCUS       V027006                 9072 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027006
VERSION     V027006
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..3752
                     /label="ATP2A3(NM_005173)"
                     /note="ATP2A3(NM_005173)"
                     /gene="ATP2A3"
     CDS             3759..3824
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3848..4436
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4502..4550
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4752..5180
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5194..5523
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5590..6384
                     /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    6560..6681
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6730..6746)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6754..6770
                     /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(6778..6808)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6823..6844
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7132..7720)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7891..8751)
                     /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(8752..8856)
                     /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 ccaccatgga ggcggcgcat ctgctcccgg
      781 ccgccgacgt gctgcgccac ttctcggtga cagccgaggg cggcctgagc ccggcgcagg
      841 tgaccggcgc gcgggagcgc tacggcccca acgagctccc gagtgaggaa gggaagtccc
      901 tgtgggagct ggtgctggaa cagtttgagg acctcctggt gcgcatcctg ctgctggctg
      961 cccttgtctc ctttgtcctg gcctggttcg aggagggcga ggagaccacg accgccttcg
     1021 tggagcccct ggtcatcatg ctgatcctcg tggccaacgc cattgtgggc gtgtggcagg
     1081 aacgcaacgc cgagagtgcc atcgaggccc tgaaggagta tgagcctgag atgggcaagg
     1141 tgatccgctc ggaccgcaag ggcgtgcaga ggatccgtgc ccgggacatc gtcccagggg
     1201 acattgtaga agtggcagtg ggggacaaag tgcctgctga cctccgcctc atcgagatca
     1261 agtccaccac gctgcgagtg gaccagtcca tcctgacggg tgaatctgtg tccgtgacca
     1321 agcacacaga ggccatccca gaccccagag ctgtgaacca ggacaagaag aacatgctgt
     1381 tttctggcac caatatcaca tcgggcaaag cggtgggtgt ggccgtggcc accggcctgc
     1441 acacggagct gggcaagatc cggagccaga tggcggcagt cgagcccgag cggacgccgc
     1501 tgcagcgcaa gctggacgag tttggacggc agctgtccca cgccatctct gtgatctgcg
     1561 tggccgtgtg ggtcatcaac atcggccact tcgccgaccc ggcccacggt ggctcctggc
     1621 tgcgtggcgc tgtctactac ttcaagatcg ccgtggccct ggcggtggcg gccatccccg
     1681 agggcctccc ggctgtcatc actacatgcc tggcactggg cacgcggcgc atggcacgca
     1741 agaacgccat cgtgcgaagc ctgccgtccg tggagaccct gggctgcacc tcagtcatct
     1801 gctccgacaa gacgggcacg ctcaccacca atcagatgtc tgtctgccgg atgttcgtgg
     1861 tagccgaggc cgatgcgggc tcctgccttt tgcacgagtt caccatctcg ggtaccacgt
     1921 atacccccga gggcgaagtg cggcaggggg atcagcctgt gcgctgcggc cagttcgacg
     1981 ggctggtgga gctggcgacc atctgcgccc tgtgcaacga ctcggctctg gactacaacg
     2041 aggccaaggg tgtgtatgag aaggtgggag aggccacgga gacagctctg acttgcctgg
     2101 tggagaagat gaacgtgttc gacaccgacc tgcaggctct gtcccgggtg gagcgagctg
     2161 gcgcctgtaa cacggtcatc aagcagctga tgcggaagga gttcaccctg gagttctccc
     2221 gagaccggaa atccatgtcc gtgtactgca cgcccacccg ccctcaccct actggccagg
     2281 gcagcaagat gtttgtgaag ggggctcctg agagtgtgat cgagcgctgt agctcagtcc
     2341 gcgtggggag ccgcacagca cccctgaccc ccacctccag ggagcagatc ctggcaaaga
     2401 tccgggattg gggctcaggc tcagacacgc tgcgctgcct ggcactggcc acccgggacg
     2461 cgcccccaag gaaggaggac atggagctgg acgactgcag caagtttgtg cagtacgaga
     2521 cggacctgac cttcgtgggc tgcgtaggca tgctggaccc gccgcgacct gaggtggctg
     2581 cctgcatcac acgctgctac caggcgggca tccgcgtggt catgatcacg ggggataaca
     2641 aaggcactgc cgtggccatc tgccgcaggc ttggcatctt tggggacacg gaagacgtgg
     2701 cgggcaaggc ctacacgggc cgcgagtttg atgacctcag ccccgagcag cagcgccagg
     2761 cctgccgcac cgcccgctgc ttcgcccgcg tggagcccgc acacaagtcc cgcatcgtgg
     2821 agaacctgca gtcctttaac gagatcactg ctatgactgg cgatggagtg aacgacgcac
     2881 cagccctgaa gaaagcagag atcggcatcg ccatgggctc aggcacggcc gtggccaagt
     2941 cggcggcaga gatggtgctg tcagatgaca actttgcctc catcgtggct gcggtggagg
     3001 agggccgggc catctacagc aacatgaagc aattcatccg ctacctcatc tcctccaatg
     3061 ttggcgaggt cgtctgcatc ttcctcacgg caattctggg cctgcccgaa gccctgatcc
     3121 ctgtgcagct gctctgggtg aacctggtga cagacggcct acctgccacg gctctgggct
     3181 tcaacccgcc agacctggac atcatggaga agctgccccg gagcccccga gaagccctca
     3241 tcagtggctg gctcttcttc cgatacctgg ctatcggagt gtacgtaggc ctggccacag
     3301 tggctgccgc cacctggtgg tttgtgtatg acgccgaggg acctcacatc aacttctacc
     3361 agctgaggaa cttcctgaag tgctccgaag acaacccgct ctttgccggc atcgactgtg
     3421 aggtgttcga gtcacgcttc cccaccacca tggccttgtc cgtgctcgtg accattgaaa
     3481 tgtgcaatgc cctcaacagc gtctcggaga accagtcgct gctgcggatg ccgccctgga
     3541 tgaacccctg gctgctggtg gctgtggcca tgtccatggc cctgcacttc ctcatcctgc
     3601 tcgtgccgcc cctgcctctc attttccagg tgaccccact gagcgggcgc cagtgggtgg
     3661 tggtgctcca gatatctctg cctgtcatcc tgctggatga ggccctcaag tacctgtccc
     3721 ggaaccacat gcacgaagaa atgagccaga agggatccga ctacaaagac catgacggtg
     3781 attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata
     3841 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3901 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3961 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4021 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4081 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4141 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4201 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     4261 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4321 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4381 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     4441 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     4501 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     4561 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     4621 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     4681 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     4741 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4801 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4861 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     4921 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4981 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     5041 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     5101 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     5161 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     5221 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     5281 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5341 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5401 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5461 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5521 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     5581 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     5641 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     5701 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5761 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5821 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5881 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5941 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     6001 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     6061 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     6121 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     6181 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     6241 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     6301 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     6361 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     6421 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     6481 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     6541 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     6601 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6661 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     6721 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6781 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6841 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6901 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6961 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     7021 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     7081 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7141 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7201 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7261 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7321 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7381 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7441 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7501 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7561 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7621 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7681 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7741 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7801 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7861 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7921 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7981 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8041 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8101 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8161 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8221 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8281 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8341 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8401 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8461 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8521 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8581 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8641 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8701 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8761 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8821 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8881 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     8941 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     9001 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     9061 aatctgctta gg
//
