LOCUS       V022897                 6881 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022897
VERSION     V022897
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..2371
                     /label="TMCC3(NM_020698)"
                     /note="TMCC3(NM_020698)"
                     /gene="TMCC3"
     CDS             2378..2443
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2481..2705
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2751..3179
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3193..3522
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3589..4383
                     /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    4557..4678
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4727..4743)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4751..4767
                     /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(4775..4805)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4820..4841
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5129..5714)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5885..6745)
                     /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(6746..6850)
                     /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 atgccgggca gcgacacggc
      961 gctcaccgtg gaccggacct actcgtaccc cggccggcac caccgctgca agagccgggt
     1021 agaacgtcat gacatgaata ccttaagcct gcccctgaac atacgccgag gggggtcaga
     1081 caccaacctc aactttgatg tcccggatgg catcctggac ttccacaagg tcaaactcac
     1141 tgcagacagc ctgaagcaaa aaattctaaa ggtaacagag cagataaaaa ttgagcaaac
     1201 atcgcgcgat gggaatgttg cggagtatct gaaactagtg aacaacgcag acaagcagca
     1261 ggcgggacgt atcaagcaag tctttgagaa gaagaatcag aaatcagctc actccatcgc
     1321 ccagctgcag aagaagttag agcagtatca tcgaaagctc agagagatcg agcagaatgg
     1381 agcctctagg agctcaaagg acatttccaa agaccacctg aaggatatac atcgctcttt
     1441 gaaagatgcc cacgtgaaat ctcgaactgc cccccattgc atggagagca gcaaatcggg
     1501 catgccaggg gtctcactta ctccacctgt gttcgttttc aataagtcca gagagtttgc
     1561 caacctgatc cggaataagt ttggcagcgc cgacaacatt gctcacttga aaaattcctt
     1621 agaggagttt aggccagagg cgagtgccag ggcctacggg ggcagcgcta ccatcgtgaa
     1681 caaacccaag tatggcagtg atgatgaatg ttcgagtggc acgtcaggct cggccgacag
     1741 taacggaaac cagtcgtttg gggctggtgg agccagcaca ctggacagcc agggcaagct
     1801 cgccgtgatc ctggaggaac tgagggagat caaggatacc caagctcagc tggctgagga
     1861 catcgaggca ctgaaggtgc agtttaagag agaatatggt tttatttctc agaccctgca
     1921 agaggaaaga tacaggtatg agcgactgga ggaccagctg catgacctga cggacctgca
     1981 tcagcatgag acagccaacc tgaagcagga gctggccagc attgaggaga aggtggccta
     2041 ccaggcctac gagcgctcgc gggacatcca ggaagccttg gaatcctgcc agactcgcat
     2101 ttctaagctg gagctccacc agcaagagca gcaagctctg cagacagaca ccgtgaatgc
     2161 taaagttctc ctggggaggt gcatcaacgt gatcctggcc ttcatgactg tcatcttagt
     2221 gtgtgtgtcc accatcgcga agttcgtctc acccatgatg aagagtcgct gccacattct
     2281 tggcaccttc tttgccgtga ctcttcttgc tatattttgt aaaaactggg accatatcct
     2341 gtgtgccata gaaaggatga taataccaag actcgaggac tacaaagacc atgacggtga
     2401 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgttta
     2461 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2521 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2581 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2641 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     2701 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     2761 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2821 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2881 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2941 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3001 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3061 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3121 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3181 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3241 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3301 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3361 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3421 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3481 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3541 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3601 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3661 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3721 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3781 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3841 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3901 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3961 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4021 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4081 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4141 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4201 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4261 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4321 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4381 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4441 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4501 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4561 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4621 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4681 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4741 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4801 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4861 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4921 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4981 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5041 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5101 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5161 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5221 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5281 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5341 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5401 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5461 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5521 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5581 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5641 gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5701 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5761 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5821 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5881 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5941 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6001 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6061 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6121 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6181 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6241 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6301 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6361 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6421 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6481 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6541 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6601 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6661 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6721 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6781 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6841 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
