LOCUS       V023294                 7597 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023294
VERSION     V023294
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             1009..3117
                     /label="USHBP1(NM_001321417)"
                     /note="USHBP1(NM_001321417)"
                     /gene="USHBP1"
     polyA_signal    3197..3421
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3467..3895
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3909..4238
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4305..5099
                     /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    5273..5394
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5443..5459)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5467..5483
                     /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(5491..5521)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5536..5557
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5845..6430)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6601..7461)
                     /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(7462..7566)
                     /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 ggtaccgagc tcggatccat gagtgcccgg
     1021 gccacgcggc cccgaagccg gcgagggagg catgctccac ccggtgaact ggatcccgtg
     1081 gctgagagtt cagaggaggt cgaggcagcc agtgggagct ccaagcccag ctttgcccca
     1141 cctccggtga gctccgggct ggagcagctg ggccccatgg aggaggtcag tggccaaggc
     1201 ctaggaagca ggactgacaa gaagatggat gggggctctg gcagggaact ggcctcagcc
     1261 cccgaagtgc cccacaaacc tgcagtggaa gcccaccaag ccccagaagc agccctacag
     1321 tacaaggaga ctgtgccccc tgggaatggg gcccccgatg tgtttcagac cctccagcac
     1381 actctgagct ccctggaggc agcggctgca gcctggcgcc accagccccc cagccattct
     1441 gggccgatgg agttcgaagg cacaagcgaa gggggagcag gttcccttgg gaagcaggaa
     1501 ggggcaggga gctgccagcg agaggcagct cgcctggccg agaggaatgc ctggctccgg
     1561 ctggccctga gtagccgaga ggatgagctg gtccgcacgc aggcctccct ggaggccatc
     1621 cgagctgaga aggagacgct gcagaaagag gttcaagagc tgcaggattc cctcttgagg
     1681 ctggagccct gcccacatct ttcccacaac caagcaggtg gctcaggcag tggttctagc
     1741 agctctgagg cagacaggga accctgggag actcaggact ccttctccct ggctcacccc
     1801 ctgcttcggc gcctccgcag ccattccagc acccagatcc ttgggtctct tcccaaccag
     1861 cccctcagtc ctgagatgca catcatggaa gcccagatgg agcaactccg ggggagcatt
     1921 gagaagctca aatgctttaa tcgtctgcta tcagctgtgc tacagggata caagggccgc
     1981 tgtgaaggcc tcagcatgca gctaggccag cgggaggctg aggccacagc attgcatctg
     2041 gccttgcagt acagtgaaca ctgtgaagag gcatacaggg ttctgcttgc tctgcgggag
     2101 gccgactcag gagcaggaga cgaagccccc atgagtgacc tgcaagcagc tgaaaaggaa
     2161 gcatggaggc tgctggcaca agaggaggct gccatggatg caggagcaca gcagaatcca
     2221 cagccaagcc ctgaaggcag cagtgtggat aagcccaccc cacaggaagt ggctttccag
     2281 ctccgaagct atgtccagcg tctccaggag cgccgttctc taatgaagat tctctcagag
     2341 cctggcccca ccttggcacc catgcccact gtgccccgtg cagaagccat ggtgcaggcc
     2401 attctgggga cccaggctgg cccagctctt ccccgactgg agaagacaca aattcagcag
     2461 gacctggtgg ccgcaaggga ggccctggcg gacctgatgc ttcggctgca gctggtgcgg
     2521 cgtgagaagc ggggcctaga gctgcgggag gctgccctcc gagccctggg tccagctcac
     2581 gtgctcctgc tggagcagct gcggtgggaa cgggcagagc tccaggctgg tggggcaaac
     2641 agcagtggcg gacatagcag cggaggtggc agcagcgggg acgaggaaga gtggtatcag
     2701 ggccttcctg ctgtccctgg tggcaccagt ggcattgatg gtggccaggt gggcagagcc
     2761 tgggacccag agaagttagc ccaggaactg gcagcatcgc tcaccaggac actggacctg
     2821 caggagcagc tgcagtctct gcgcagggag ctggaacagg tggctcagaa ggggcgagcc
     2881 agacggtctc agagtgccga gctgaacagg gatttatgca aagcccacag cgccctggtc
     2941 ctggccttcc gaggagccca ccggaagcag gaagagcaac gccggaagtt ggagcagcag
     3001 atggcactca tggaggctca gcaggccgag gaggtggcgg tgctcgaagc cactgcaagg
     3061 gccctgggga agcccaggcc tcccctcccg cctccccagc ttggggacac ctttctgtag
     3121 gaattctgca gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc
     3181 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3241 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3301 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3361 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3421 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3481 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3541 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3601 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3661 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3721 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3781 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3841 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3901 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3961 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4021 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4081 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4141 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4201 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4261 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4321 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4381 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4441 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4501 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4561 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4621 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4681 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4741 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4801 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4861 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4921 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4981 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5041 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5101 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5161 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5221 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5281 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5341 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5401 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5461 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5521 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5581 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5641 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5701 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5761 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5821 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5881 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5941 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6001 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6061 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6121 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6181 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6241 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6301 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6361 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6421 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6481 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6541 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6601 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6661 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6721 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6781 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6841 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6901 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6961 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7021 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7081 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7141 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7201 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7261 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7321 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7381 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7441 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7501 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7561 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
