LOCUS       V037993                 8044 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037993
VERSION     V037993
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..3564
                     /label="TTC7A(NM_020458)"
                     /note="TTC7A(NM_020458)"
                     /gene="TTC7A"
     polyA_signal    3644..3868
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3914..4342
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4356..4685
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4752..5546
                     /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    5720..5841
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5890..5906)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5914..5930
                     /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(5938..5968)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5983..6004
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6292..6877)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7048..7908)
                     /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(7909..8013)
                     /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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atggctgcga agggcgcgca cggctcctac
     1021 ctgaaggtgg agagcgagct ggagcgctgc cgcgccgagg gccactggga ccgcatgccg
     1081 gagctggtcc ggcagctgca gacgctgagc atgcccggcg gcggaggtaa caggcgaggc
     1141 agcccgagcg cagcgttcac ctttccggac accgatgact ttgggaaatt gctgctggct
     1201 gaggccctcc tggagcagtg tttgaaggag aaccatgcca aaataaaaga ctccatgcct
     1261 ttgctggaga agaatgagcc gaagatgagc gaagccaaaa attatctaag cagtatcctt
     1321 aaccatggga ggctctcgcc acagtacatg tgtgaggcca tgctgatcct gggcaaactg
     1381 cattacgtgg agggctcata ccgagatgcc atcagcatgt acgcacgggc cgggattgat
     1441 gacatgtcca tggagaacaa gcccctgtat cagatgcggc tgctgtcgga ggcttttgtc
     1501 atcaaaggcc tctctctgga acgcctaccc aactccatcg cctcccgctt ccgcctgaca
     1561 gagagggagg aggaagtgat cacctgtttt gagagggcct cctggatcgc tcaggtgttc
     1621 ctgcaggaat tggagaagac cacaaataac agcacgtcga ggcatctgaa aggctgtcac
     1681 ccgcttgact atgagctcac ctacttcctg gaagctgccc tccagagcgc ctatgtgaaa
     1741 aacctgaaga aggggaacat cgtgaagggc atgagagagc tccgggaggt gctgcggact
     1801 gtggagacca aagcaactca gaacttcaaa gtgatggcgg ccaagcacct ggcgggggtc
     1861 ctgctgcact ccctgagtga ggagtgctac tggagccccc tgtcccaccc tctgcctgag
     1921 ttcatgggca aggaggagag ttctttcgcc actcaggccc tgcggaaacc tcacctctat
     1981 gaaggagaca acctctactg ccccaaggac aacatcgagg aagccctcct gctcctcctc
     2041 atcagcgaat ccatggcaac tcgagatgtg gtgctgagcc gggtgccgga gcaggaggag
     2101 gaccggacag tgagcttgca gaatgccgca gccatctatg acctcctgag catcacgttg
     2161 ggcagaaggg gacagtacgt catgctctcg gagtgcctgg agcgagccat gaagtttgcg
     2221 tttggagaat ttcacctttg gtaccaggtg gccctctcca tggtggcttg tgggaagtca
     2281 gcctacgctg tgtccctgct gcgggagtgt gtgaagttgc ggccctcgga ccccaccgtg
     2341 cccctgatgg ccgcgaaggt ctgcatcggg tcccttcgct ggctagagga agcagagcac
     2401 tttgccatga tggtgatcag cctcggagag gaagccgggg agttcctccc caagggctac
     2461 ctggctctgg gtctcaccta tagcctgcag gccaccgacg ccaccctgaa gtccaagcaa
     2521 gatgaattgc accggaaggc actgcagacg ctggagaggg ctcagcagct ggcgcccagt
     2581 gacccccagg tcatcctcta tgtctcgctg cagctggccc tcgtccgaca gatctccagt
     2641 gccatggagc agctgcagga ggccctgaag gtacgcaagg atgatgccca cgccctccac
     2701 ctgctggcac tgctcttctc tgcccagaag caccaccagc atgccctgga tgttgtcaac
     2761 atggccatca ccgagcaccc tgagaacttc aacctgatgt tcaccaaggt gaagctggag
     2821 caggtgctga aaggcccaga ggaagccctc gtgacctgca gacaagtgct gaggctgtgg
     2881 cagaccctgt acagcttctc ccagctggga ggcctagaaa aggatggcag cttcggtgag
     2941 ggcctcacca tgaagaagca gagtggcatg cacctgactt tgcctgatgc ccatgatgca
     3001 gactctggct cccggcgggc ttcgtccatc gccgcctccc ggctggagga ggccatgtca
     3061 gagctgacta tgccctcttc ggtcctgaag cagggcccca tgcagctgtg gaccacgctg
     3121 gaacagatct ggctgcaggc tgctgagctg ttcatggagc agcagcacct caaggaagca
     3181 ggtttctgca tccaggaggc ggcgggcctc ttccccactt ctcactcagt actctatatg
     3241 cggggccggc tggctgaggt gaagggcaac ctggaggagg ccaagcagct gtacaaggag
     3301 gcgctcacgg tgaacccaga tggcgtgcgc atcatgcata gcctgggtct gatgctgagt
     3361 cggctgggcc acaagagctt ggcccagaag gtgcttcgtg atgccgtgga gaggcagagt
     3421 acgtgccacg aggcgtggca gggcctgggc gaggtgctgc aggcccaggg ccagaacgag
     3481 gctgccgttg actgcttcct caccgccctt gagctggagg ccagcagccc tgtactgccc
     3541 ttctccatca tccccagaga gctctgagaa ttctgcagat atccagcaca gtggcggccg
     3601 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     3661 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3721 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3781 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3841 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3901 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3961 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4021 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4081 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4141 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4201 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4261 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4321 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4381 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4441 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4501 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4561 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4621 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4681 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4741 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4801 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4861 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4921 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4981 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     5041 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5101 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5161 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5221 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     5281 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5341 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5401 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5461 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5521 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5581 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5641 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5701 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5761 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5821 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5881 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5941 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6001 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6061 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6121 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6181 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6241 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6301 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6361 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6421 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6481 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6541 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6601 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6661 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6721 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6781 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg
     6841 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6901 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6961 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7021 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7081 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7141 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7201 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7261 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7321 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7381 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7441 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7501 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7561 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7621 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7681 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7741 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7801 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7861 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7921 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7981 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8041 cgtc
//
