LOCUS       V024342                 6791 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024342
VERSION     V024342
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..2281
                     /label="Tekt4(NM_027951)"
                     /note="Tekt4(NM_027951)"
                     /gene="Tekt4"
     CDS             2288..2353
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2391..2615
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2661..3089
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3103..3432
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3499..4293
                     /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    4467..4588
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4637..4653)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4661..4677
                     /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(4685..4715)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4730..4751
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5039..5624)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5795..6655)
                     /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(6656..6760)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggcccaaa cgggcgtgct
      961 cctgaccaag gagccggccc cacagtcaat agatgtctgt gagcttccgc gcaaagaata
     1021 tgaagtggcc tgcaatactg gcgcctacac gtcctcgggt ctggccaccg ctggcttccg
     1081 cacagccaaa tacctgaggg acgagtggtt tcagaactct tatgcccgct accaccaggc
     1141 cttcgcagac cgggactact ctgagcggca gcggcatgag agcgggcagc tggtagccga
     1201 gactggggca ctggcccagc gcacacaact tgactccaca agaaaggtgg gagagcgcct
     1261 ggaggacatg cattgctgga agtcagagct gcagcgagag atagacgagc tgtcttccga
     1321 gacggacctg atgatggccc agaagctgcg gctacagcgc gccttggatg ccacctccgt
     1381 gccttactcc attgctactg acaacctgca gtgcagagaa cgccgccagc acccagacct
     1441 tgtccgggac tatgtggagg tggagctact gaaggaaacc gagctgatca ggaacatcca
     1501 ggaactcctg aagaggacga taggacaagc cgtggaccag atacggctga acagggagca
     1561 taaggagtcg tgtgagatga actggtcgga caaagtggaa gtctacaaca ttgacgatac
     1621 ctgctcccgc tacaccaatg agagcaccca ggtgcagttc tatccgcatt ccagcaagtt
     1681 cgaggagagt gcttccacgc ccgagacgtg ggccaagttc aatcacgaca acctgcttcg
     1741 agccgagcgc gagcggctag cctcggttaa cctgcggaag ttaattgact gcatcttgcg
     1801 ggatacggcc gaggacctgc ggctacagtg cgatgcagtg aactcggctt tctcaagccg
     1861 ctgccaagag ctggacgact cactccagaa gctgcagtac caccttcgaa agaccctgac
     1921 ggagatcaca gaccaggaac accagattgc agccttgaag caggccatca aggacaagga
     1981 ggcccctctg cgtgtggccc agactcgcct ttaccagcgg tcacaccggc ccaatgtgga
     2041 gttgtgccgg gacaatgccc agttcaggtt gctgagcgag gtggaggagt tgaacatgtc
     2101 cctcagagcg ttaaaggaga agcttcagga tgcggagcag gcactgcgga acctggagga
     2161 ttcacgcatg agtctggaga aggacatcgc agtcaagacc aacagcctct ttatcgaccg
     2221 ccagaagtgt atgacccacc gcaaccgcta ccccagtgtc ctccagctgg ctggctacca
     2281 gctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     2341 tgacgatgac aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     2401 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2461 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2521 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2581 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2641 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2701 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2761 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2821 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2881 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2941 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3001 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3061 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3121 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3181 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3241 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3301 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3361 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3421 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     3481 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3541 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3601 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3661 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3721 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3781 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3841 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3901 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3961 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4021 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4081 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4141 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4201 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4261 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4321 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4381 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4441 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4501 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4561 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     4621 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4681 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4741 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4801 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4861 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4921 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4981 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5041 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5101 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5161 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5221 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5281 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5341 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5401 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5461 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5521 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt
     5581 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5641 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5701 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5761 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5821 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5881 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5941 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6001 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6061 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6121 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6181 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6241 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6301 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6361 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6421 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6481 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6541 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6601 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6661 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6721 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6781 cacctgacgt c
//
