LOCUS       V036612                 7237 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036612
VERSION     V036612
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             706..1893
                     /label="CTSE(NM_001910)"
                     /note="CTSE(NM_001910)"
                     /gene="CTSE"
     CDS             1900..1989
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2013..2601
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2667..2715
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2917..3345
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3359..3688
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3755..4549
                     /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    4725..4846
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4895..4911)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4919..4935
                     /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(4943..4973)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4988..5009
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5297..5885)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6056..6916)
                     /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(6917..7021)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgaa aacgctcctt
      721 cttttgctgc tggtgctcct ggagctggga gaggcccaag gatcccttca cagggtgccc
      781 ctcaggaggc atccgtccct caagaagaag ctgcgggcac ggagccagct ctctgagttc
      841 tggaaatccc ataatttgga catgatccag ttcaccgagt cctgctcaat ggaccagagt
      901 gccaaggaac ccctcatcaa ctacttggat atggaatact tcggcactat ctccattggc
      961 tccccaccac agaacttcac tgtcatcttc gacactggct cctccaacct ctgggtcccc
     1021 tctgtgtact gcactagccc agcctgcaag acgcacagca ggttccagcc ttcccagtcc
     1081 agcacataca gccagccagg tcaatctttc tccattcagt atggaaccgg gagcttgtcc
     1141 gggatcattg gagccgacca agtctctgtg gaaggactaa ccgtggttgg ccagcagttt
     1201 ggagaaagtg tcacagagcc aggccagacc tttgtggatg cagagtttga tggaattctg
     1261 ggcctgggat acccctcctt ggctgtggga ggagtgactc cagtatttga caacatgatg
     1321 gctcagaacc tggtggactt gccgatgttt tctgtctaca tgagcagtaa cccagaaggt
     1381 ggtgcgggga gcgagctgat ttttggaggc tacgaccact cccatttctc tgggagcctg
     1441 aattgggtcc cagtcaccaa gcaagcttac tggcagattg cactggataa catccaggtg
     1501 ggaggcactg ttatgttctg ctccgagggc tgccaggcca ttgtggacac agggacttcc
     1561 ctcatcactg gcccttccga caagattaag cagctgcaaa acgccattgg ggcagccccc
     1621 gtggatggag aatatgctgt ggagtgtgcc aaccttaacg tcatgccgga tgtcaccttc
     1681 accattaacg gagtccccta taccctcagc ccaactgcct acaccctact ggacttcgtg
     1741 gatggaatgc agttctgcag cagtggcttt caaggacttg acatccaccc tccagctggg
     1801 cccctctgga tcctggggga tgtcttcatt cgacagtttt actcagtctt tgaccgtggg
     1861 aataaccgtg tgggactggc cccagcagtc cccctcgagg aacaaaaact aatatcagag
     1921 gaagatttgg aacagaaact aatcagtgaa gaagatttag agcaaaagtt aatttccgag
     1981 gaggacttat agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg
     2041 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     2101 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     2161 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     2221 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     2281 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     2341 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     2401 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     2461 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     2521 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     2581 ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac
     2641 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg
     2701 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     2761 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     2821 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     2881 cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     2941 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3001 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3061 ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3121 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3181 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3241 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct
     3301 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     3361 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3421 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3481 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3541 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3601 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     3661 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     3721 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     3781 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     3841 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     3901 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     3961 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4021 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4081 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4141 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4201 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4261 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4321 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4381 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4441 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4501 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4561 gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4621 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4681 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     4741 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4801 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     4861 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4921 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4981 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5041 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5101 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5161 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5221 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5281 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5341 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5401 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5461 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     5521 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5581 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5641 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5701 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     5761 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5821 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5881 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5941 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6001 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6061 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6121 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6181 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6241 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6301 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6361 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6421 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6481 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6541 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6601 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6661 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6721 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6781 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6841 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6901 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6961 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7021 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     7081 atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     7141 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     7201 aggcttgacc gacaattgca tgaagaatct gcttagg
//
