LOCUS       V039629                 6487 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039629
VERSION     V039629
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..2007
                     /label="CTSB(NM_001908)"
                     /note="CTSB(NM_001908)"
                     /gene="CTSB"
     polyA_signal    2087..2311
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2357..2785
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2799..3128
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3195..3989
                     /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    4163..4284
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4333..4349)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4357..4373
                     /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(4381..4411)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4426..4447
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4735..5320)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5491..6351)
                     /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(6352..6456)
                     /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 atgtggcagc tctgggcctc cctctgctgc
     1021 ctgctggtgt tggccaatgc ccggagcagg ccctctttcc atcccctgtc ggatgagctg
     1081 gtcaactatg tcaacaaacg gaataccacg tggcaggccg ggcacaactt ctacaacgtg
     1141 gacatgagct acttgaagag gctatgtggt accttcctgg gtgggcccaa gccaccccag
     1201 agagttatgt ttaccgagga cctgaagctg cctgcaagct tcgatgcacg ggaacaatgg
     1261 ccacagtgtc ccaccatcaa agagatcaga gaccagggct cctgtggctc ctgctgggcc
     1321 ttcggggctg tggaagccat ctctgaccgg atctgcatcc acaccaatgc gcacgtcagc
     1381 gtggaggtgt cggcggagga cctgctcaca tgctgtggca gcatgtgtgg ggacggctgt
     1441 aatggtggct atcctgctga agcttggaac ttctggacaa gaaaaggcct ggtttctggt
     1501 ggcctctatg aatcccatgt agggtgcaga ccgtactcca tccctccctg tgagcaccac
     1561 gtcaacggct cccggccccc atgcacgggg gagggagata cccccaagtg tagcaagatc
     1621 tgtgagcctg gctacagccc gacctacaaa caggacaagc actacggata caattcctac
     1681 agcgtctcca atagcgagaa ggacatcatg gccgagatct acaaaaacgg ccccgtggag
     1741 ggagctttct ctgtgtattc ggacttcctg ctctacaagt caggagtgta ccaacacgtc
     1801 accggagaga tgatgggtgg ccatgccatc cgcatcctgg gctggggagt ggagaatggc
     1861 acaccctact ggctggttgc caactcctgg aacactgact ggggtgacaa tggcttcttt
     1921 aaaatactca gaggacagga tcactgtgga atcgaatcag aagtggtggc tggaattcca
     1981 cgcaccgatc agtactggga aaagatctaa gaattctgca gatatccagc acagtggcgg
     2041 ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     2101 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2161 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2221 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2281 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     2341 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2401 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     2461 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2521 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     2581 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2641 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     2701 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     2761 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     2821 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2881 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2941 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3001 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3061 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3121 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     3181 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     3241 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     3301 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     3361 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     3421 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     3481 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     3541 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     3601 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     3661 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     3721 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     3781 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     3841 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     3901 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     3961 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     4021 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     4081 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     4141 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     4201 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4261 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     4321 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4381 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4441 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4501 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4561 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4621 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4681 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4741 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4801 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4861 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4921 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4981 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5041 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5101 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5161 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5221 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     5281 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5341 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5401 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5461 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5521 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5581 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5641 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5701 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5761 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5821 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5881 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5941 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6001 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6061 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6121 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6181 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6241 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6301 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6361 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6421 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6481 tgacgtc
//
