LOCUS       V047173                 8892 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047173
VERSION     V047173
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..3572
                     /label="Trak1(NM_175114)"
                     /note="Trak1(NM_175114)"
                     /gene="Trak1"
     CDS             3579..3644
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3668..4256
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4322..4370
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4572..5000
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5014..5343
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5410..6204
                     /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    6380..6501
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6550..6566)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6574..6590
                     /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(6598..6628)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6643..6664
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6952..7540)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7711..8571)
                     /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(8572..8676)
                     /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 ccaccatggc attggcaatt cagctcaggc
      781 agccctccag ggcccaacct ctgccaggac tcagccacac actggctggg acagactcat
      841 gtgacgtgtg caacagcacc aaccttccag aagttgagat cattagcctg ctggaggaac
      901 agctgcccca ttacaagcta agagcggaca ccatctatgg ttacgaccac gatgactggc
      961 tgcatacgcc cctcatctcc ccagatgcca acatcgacct cacaactgag cagatcgaag
     1021 agacgctgaa atacttcctt ctatgtgccg aaagagttgg ccagatgact aagacataca
     1081 atgacattga tgctgtcacg aggcttcttg aggagaaaga gcgggatttg gagctggctg
     1141 cgaggatcgg tcagtcattg ttgaagaaga acaagaccct aactgagagg aatgaactgc
     1201 tggaagagca ggtggagcac atccgggagg aggtgtctca gctccgacat gagctgtcca
     1261 tgaaagacga gctgcttcaa ttctacacca gtgccgctga ggagagcgag cctgagtccg
     1321 tctgctcaac cccgctgaag aggaacgagt cctcctcctc cgtccagaat tacttccacc
     1381 tggactctct tcagaagaag ctcaaggacc tcgaagagga gaacgttgta cttcgatccg
     1441 aggcctgtca gctgaagaca gagaccatca cctatgagga gaaggagcag cagctggtga
     1501 acgactgcgt gaaggagctg agggatgcca acgtccagat tgcgagcatc tctgaggagc
     1561 tggctaagaa gacggaagat gccgcccgcc agcaggagga gatcacccac ctgctgtcgc
     1621 aaatcgtgga cttacagaag aaggcaaaat cttgcgcggt ggaaaatgaa gagcttgtcc
     1681 agcacctggg cgctgccaag gacgcccagc gtcaactcac agccgagctc cgagagctgg
     1741 aggataagta cgcggagtgc atggagatgc ttcatgaggc tcaggaggag ttgaagaacc
     1801 tgcggaacaa gacgatgccc acgtcccggc gctaccactc tctgggcctg ttccccatgg
     1861 actcgttggc tgcagaaatc gagggaacta tgcgcaagga gctgcagctg gaagagctgg
     1921 agtctccaga catcacccac cagaagcgtg tcttcgagac ggtgaggaac gtcaatcaag
     1981 tggtcaagca gagatcacta accccttctc ccatgaacat cccgggctcc aaccagtcct
     2041 cagccatgaa ctccctcctg tccagctgcg tcagcacccc tcgctccagc ttctacggca
     2101 gcgacgtcag caacgtggtc ctcgataaca aaaccaacag catcctcctg gagaccgagg
     2161 cagccgacct ggggaacgag gaccacaata agaagccggg cactccaggc acgccaggct
     2221 cccatgacct ggagacggca cttaggcggc tatccctacg ccgggagaac tacctgtctg
     2281 agcggaggtt cttcgaggag gagcaggaga ggaagctgcg agagctggct gagaaggggg
     2341 agctgcacag tggctcgctc acgcccaccg agagcatcat gtccctggga acacactcac
     2401 gcttctccga gttcacgggc ttctctggca tgtccttcag cagccgctcc tacctgcccg
     2461 agaagctgca gatcgtgaag ccactcgaag gttcagccac gcttcaccac tggcagcaat
     2521 tggcccagcc tcatcttggg ggcatcctgg acccccgtcc tggtgtggtc accaagggct
     2581 tccggacttt ggatgttgac ctggatgaag tgtactgcct taacgacttt gaggaagatg
     2641 acacaggtga ccacatttct ctggcgggcc tagctacctc cacgccaatt cagcacccgg
     2701 agacctcagc gcaccaccct gggaagtgca tgtcacagac caactcaact ttcaccttta
     2761 ccacttgccg catcctgcac ccttcagacg agctcactcg ggtcacgcca agccttaact
     2821 ctgccccagc tccggcttgt agcagtacca gccacttgaa atccacacca gtggccacgc
     2881 catgcacccc ccggagactg agcctggctg agtctttcac aaatgtccgt gagtccacga
     2941 caaccatgag cacgtccctg gggctggtat ggttgctgaa ggagcgaggc atctcggcgg
     3001 ccgtgtacga cccccagagt tgggacagag ccggcagggg ctccctcctc cactcctaca
     3061 cccctaggat ggctgtgatc ccgtccaccc caccaaactc ccctatgcag acaccttcgg
     3121 cctccccgcc ctccttcgag ttcaagtgca cgagccctcc ctacaacaac ttcctggctt
     3181 ccaagccggc cagctctatc ctgagggagg tgagggaaaa gaggcccgtg cggagcagcg
     3241 agagccaaac agatgtgtct gtctccaacc tcaaccttgt ggacaaagtc aggaggtttg
     3301 gggtggccag agtggtgaac tcaggacgtg cccgtatccc caccttgact gaggaacagg
     3361 gacctctcct ctgtggaccc acagggccag cacaggctct tgtccctggg ggcctggtcc
     3421 ccgagggcct gcctctgggg tgccccagtg gcatccgacg gaatcgcagc ttccccacta
     3481 tggtcgggtc cagcgtgcag atgagagccc ccgtgatcct cacctcgggc atcttgatgg
     3541 gtgctaagct ccccaaacag actagcttgc ggggatccga ctacaaagac catgacggtg
     3601 attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata
     3661 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3721 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3781 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3841 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3901 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3961 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4021 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     4081 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4141 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4201 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     4261 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     4321 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     4381 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     4441 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     4501 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     4561 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4621 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4681 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     4741 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4801 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4861 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4921 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     4981 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     5041 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     5101 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5161 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5221 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5281 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5341 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     5401 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     5461 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     5521 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5581 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5641 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5701 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5761 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5821 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5881 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5941 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     6001 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     6061 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     6121 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     6181 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     6241 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     6301 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     6361 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     6421 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6481 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     6541 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6601 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6661 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6721 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6781 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6841 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6901 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6961 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7021 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7081 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7141 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7201 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7261 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7321 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7381 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7441 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7501 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7561 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7621 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7681 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7741 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7801 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7861 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7921 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7981 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8041 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8101 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8161 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8221 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8281 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8341 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8401 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8461 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8521 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8581 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8641 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8701 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     8761 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     8821 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     8881 aatctgctta gg
//
