LOCUS       V021572                 8777 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021572
VERSION     V021572
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             683..3400
                     /label="CBL(NM_005188)"
                     /note="CBL(NM_005188)"
                     /gene="CBL"
     CDS             3440..3529
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3553..4141
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4207..4255
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4457..4885
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4899..5228
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5295..6089
                     /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    6265..6386
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6435..6451)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6459..6475
                     /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(6483..6513)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6528..6549
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6837..7425)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7596..8456)
                     /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(8457..8561)
                     /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 ggatccgcca ccatggccgg caacgtgaag aagagctctg gggccggggg
      721 cggcagcggc tccgggggct cgggttcggg tggcctgatt gggctcatga aggacgcctt
      781 ccagccgcac caccaccacc accaccacct cagcccccac ccgccgggga cggtggacaa
      841 gaagatggtg gagaagtgct ggaagctcat ggacaaggtg gtgcggttgt gtcagaaccc
      901 aaagctggcg ctaaagaata gcccacctta tatcttagac ctgctaccag atacctacca
      961 gcatctccgt actatcttgt caagatatga ggggaagatg gagacacttg gagaaaatga
     1021 gtattttagg gtgtttatgg agaatttgat gaagaaaact aagcaaacca taagcctctt
     1081 caaggaggga aaagaaagaa tgtatgagga gaattctcag cctaggcgaa acctaaccaa
     1141 actgtccctc atcttcagcc acatgctggc agaactaaaa ggaatctttc caagtggact
     1201 ctttcaggga gacacatttc ggattactaa agcagatgct gcggaatttt ggagaaaagc
     1261 ttttggggaa aagacaatag tcccttggaa gagctttcga caggctctac atgaagtgca
     1321 tcccatcagt tctgggctgg aggccatggc tctgaaatcc actattgatc tgacctgcaa
     1381 tgattatatt tcggtttttg aatttgacat ctttacccga ctctttcagc cctggtcctc
     1441 tttgctcagg aattggaaca gccttgctgt aactcatcct ggctacatgg cttttttgac
     1501 gtatgacgaa gtgaaagctc ggctccagaa attcattcac aaacctggca gttatatctt
     1561 ccggctgagc tgtactcgtc tgggtcagtg ggctattggg tatgttactg ctgatgggaa
     1621 cattctccag acaatccctc acaataaacc tctcttccaa gcactgattg atggcttcag
     1681 ggaaggcttc tatttgtttc ctgatggacg aaatcagaat cctgatctga ctggcttatg
     1741 tgaaccaact ccccaagacc atatcaaagt gacccaggaa caatatgaat tatactgtga
     1801 gatgggctcc acattccaac tatgtaaaat atgtgctgaa aatgataagg atgtaaagat
     1861 tgagccctgt ggacacctca tgtgcacatc ctgtcttaca tcctggcagg aatcagaagg
     1921 tcagggctgt cctttctgcc gatgtgaaat taaaggtact gaacccatcg tggtagatcc
     1981 gtttgatcct agagggagtg gcagcctgtt gaggcaagga gcagagggag ctccctcccc
     2041 aaattatgat gatgatgatg atgaacgagc tgatgatact ctcttcatga tgaaggaatt
     2101 ggctggtgcc aaggtggaac ggccgccttc tccattctcc atggccccac aagcttccct
     2161 tcccccggtg ccaccacgac ttgaccttct gccgcagcga gtatgtgttc cctcaagtgc
     2221 ttctgctctt ggaactgctt ctaaggctgc ttctggctcc cttcataaag acaaaccatt
     2281 gccagtacct cccacacttc gagatcttcc accaccaccg cctccagacc ggccatattc
     2341 tgttggagca gaatcccgac ctcaaagacg ccccttgcct tgtacaccag gcgactgtcc
     2401 ctccagagac aaactgcccc ctgtcccctc tagccgcctt ggagactcat ggctgccccg
     2461 gccaatcccc aaagtaccag tatctgcccc aagttccagt gatccctgga caggaagaga
     2521 attaaccaac cggcactcac ttccattttc attgccctca caaatggagc ccagaccaga
     2581 tgtgcctagg ctcggaagca cgttcagtct ggatacctcc atgagtatga atagcagccc
     2641 attagtaggt ccagagtgtg accaccccaa aatcaaacct tcctcatctg ccaatgccat
     2701 ttattctctg gctgccagac ctcttcctgt gccaaaactg ccacctgggg agcaatgtga
     2761 gggtgaagag gacacagagt acatgactcc ctcttccagg cctctacggc ctttggatac
     2821 atcccagagt tcacgagcat gtgattgcga ccagcagatt gatagctgta cgtatgaagc
     2881 aatgtataat attcagtccc aggcgccatc tatcaccgag agcagcacct ttggtgaagg
     2941 gaatttggcc gcagcccatg ccaacactgg tcccgaggag tcagaaaatg aggatgatgg
     3001 gtatgatgtc ccaaagccac ctgtgccggc cgtgctggcc cgccgaactc tctcagatat
     3061 ctctaatgcc agctcctcct ttggctggtt gtctctggat ggtgatccta caacaaatgt
     3121 cactgaaggt tcccaagttc ccgagaggcc tccaaaacca ttcccgcgga gaatcaactc
     3181 tgaacggaaa gctggcagct gtcagcaagg tagtggtcct gccgcctctg ctgccaccgc
     3241 ctcacctcag ctctccagtg agatcgagaa cctcatgagt caggggtact cctaccagga
     3301 catccagaaa gctttggtca ttgcccagaa caacatcgag atggccaaaa acatcctccg
     3361 ggaatttgtt tccatttctt ctcctgccca tgtagctacc gaattctgca gatatccagc
     3421 acagtggcgg ccgctcgagg aacaaaaact aatatcagag gaagatttgg aacagaaact
     3481 aatcagtgaa gaagatttag agcaaaagtt aatttccgag gaggacttat agtgagggcc
     3541 cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     3601 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     3661 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     3721 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     3781 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     3841 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3901 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     3961 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     4021 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     4081 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     4141 cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     4201 ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa
     4261 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     4321 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     4381 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc
     4441 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4501 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4561 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct
     4621 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4681 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4741 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4801 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg
     4861 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4921 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4981 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5041 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5101 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5161 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5221 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     5281 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5341 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     5401 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     5461 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5521 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5581 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5641 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5701 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5761 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5821 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5881 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5941 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     6001 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     6061 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac
     6121 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     6181 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     6241 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     6301 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6361 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     6421 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6481 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6541 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6601 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6661 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6721 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6781 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6841 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6901 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6961 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     7021 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     7081 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7141 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7201 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7261 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7321 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7381 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7441 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7501 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7561 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7621 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     7681 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     7741 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7801 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7861 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7921 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7981 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     8041 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     8101 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     8161 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     8221 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     8281 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     8341 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     8401 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     8461 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     8521 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     8581 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa
     8641 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     8701 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     8761 tgaagaatct gcttagg
//
