LOCUS       V024003                 8623 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024003
VERSION     V024003
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             775..3279
                     /label="TRIM28(NM_005762)"
                     /note="TRIM28(NM_005762)"
                     /gene="TRIM28"
     CDS             3280..3297
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     misc_feature    3399..3987
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4053..4101
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4303..4731
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4745..5074
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5141..5935
                     /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    6111..6232
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6281..6297)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6305..6321
                     /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(6329..6359)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6374..6395
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6683..7271)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7442..8302)
                     /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(8303..8407)
                     /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 gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagc tagcgtttaa acttaagctt ggtaccgagc tcggatccgc caccatggcg
      781 gcctccgcgg cggcagcctc ggcagcagcg gcctcggccg cctctggcag cccgggcccg
      841 ggcgagggct ccgctggcgg cgaaaagcgc tccaccgccc cttcggccgc agcctcggcc
      901 tctgcctcag ccgcggcgtc gtcgcccgcg gggggcggcg ccgaggcgct ggagctgctg
      961 gagcactgcg gcgtgtgcag agagcgcctg cgacccgaga gggagccccg cctgctgccc
     1021 tgtttgcact cggcctgtag tgcctgctta gggcccgcgg cccccgccgc cgccaacagc
     1081 tcgggggacg gcggggcggc gggcgacggc accgtggtgg actgtcccgt gtgcaagcaa
     1141 cagtgcttct ccaaagacat cgtggagaat tatttcatgc gtgatagtgg cagcaaggct
     1201 gccaccgacg cccaggatgc gaaccagtgc tgcactagct gtgaggataa tgccccagcc
     1261 accagctact gtgtggagtg ctcggagcct ctgtgtgaga cctgtgtaga ggcgcaccag
     1321 cgggtgaagt acaccaagga ccatactgtg cgctctactg ggccagccaa gtctcgggat
     1381 ggtgaacgta ctgtctattg caacgtacac aagcatgaac cccttgtgct gttttgtgag
     1441 agctgtgata ctctcacctg ccgagactgc cagctcaatg cccacaagga ccaccagtac
     1501 cagttcttag aggatgcagt gaggaaccag cgcaagctcc tggcctcact ggtgaagcgc
     1561 cttggggaca aacatgcaac attgcagaag agcaccaagg aggttcgcag ctcaatccgc
     1621 caggtgtctg acgtacagaa gcgtgtgcaa gtggatgtca agatggccat cctgcagatc
     1681 atgaaggagc tgaataagcg gggccgtgtg ctggtcaatg atgcccagaa ggtgactgag
     1741 gggcagcagg agcgcctgga gcggcagcac tggaccatga ccaagatcca gaagcaccag
     1801 gagcacattc tgcgctttgc ctcttgggct ctggagagtg acaacaacac agcccttttg
     1861 ctttctaaga agttgatcta cttccagctg caccgggccc tcaagatgat tgtggatccc
     1921 gtggagccac atggcgagat gaagtttcag tgggacctca atgcctggac caagagtgcc
     1981 gaggcctttg gcaagattgt ggcagagcgt cctggcacta actcaacagg ccctgcaccc
     2041 atggcccctc caagagcccc agggcccctg agcaagcagg gctctggcag cagccagccc
     2101 atggaggtgc aggaaggcta tggctttggg tcaggagatg atccctactc aagtgcagag
     2161 ccccatgtgt caggtgtgaa acggtcccgc tcaggtgagg gcgaggtgag cggccttatg
     2221 cgcaaggtgc cacgagtgag ccttgaacgc ctggacctgg acctcacagc tgacagccag
     2281 ccacccgtct tcaaggtctt cccaggcagt accactgagg actacaacct tattgttatt
     2341 gaacgtggcg ctgccgctgc agctaccggc cagccaggga ctgcgcctgc aggaacccct
     2401 ggtgccccac ccctggctgg catggccatt gtcaaggagg aggagacgga ggctgccatt
     2461 ggagcccctc ctactgccac tgagggccct gagaccaaac ctgtgcttat ggctcttgcg
     2521 gagggtcctg gtgctgaggg tccccgcctg gcctcaccta gtggcagcac cagctcaggg
     2581 ctggaggtgg tggctcctga gggtacctca gccccaggtg gtggcccggg aaccctggat
     2641 gacagtgcca ccatttgccg tgtctgccag aagccaggcg atctggttat gtgcaaccag
     2701 tgtgagtttt gtttccacct ggactgtcac ctgccggccc tgcaggatgt accaggggag
     2761 gagtggagct gctcactctg ccatgtgctc cctgacctga aggaggagga tggcagcctc
     2821 agcctggatg gtgcagacag cactggcgtg gtggccaagc tctcaccagc caaccagcgg
     2881 aaatgtgagc gtgtactgct ggccctattc tgtcacgaac cctgccgccc cctgcatcag
     2941 ctggctaccg actccacctt ctccctggac cagcccggtg gcaccctgga tctgaccctg
     3001 atccgtgccc gcctccagga gaagttgtca cctccctaca gctccccaca ggagtttgcc
     3061 caggatgtgg gccgcatgtt caagcaattc aacaagttaa ctgaggacaa ggcagacgtg
     3121 cagtccatca tcggcctgca gcgcttcttc gagacgcgca tgaacgaggc cttcggtgac
     3181 accaagttct ctgctgtgct ggtggagccc ccgccgatga gcctgcctgg tgctggcctg
     3241 agttcccagg agctgtctgg tggccctggt gatggccccc accatcacca tcaccattag
     3301 aattctgcag atatccagca cagtggcggc cgctcgagtc tagagggccc cttaagggtt
     3361 cgatccctac cggttagtaa tgagtttgat atctcgacaa tcaacctctg gattacaaaa
     3421 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     3481 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     3541 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     3601 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     3661 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     3721 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     3781 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc
     3841 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     3901 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     3961 ggatctccct ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa
     4021 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     4081 cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     4141 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     4201 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     4261 ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     4321 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4381 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4441 gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg
     4501 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4561 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4621 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt
     4681 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     4741 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4801 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4861 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4921 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4981 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     5041 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     5101 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     5161 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     5221 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     5281 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     5341 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     5401 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     5461 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     5521 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     5581 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     5641 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     5701 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     5761 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     5821 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     5881 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     5941 actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     6001 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     6061 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     6121 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     6181 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     6241 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     6301 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6361 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6421 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6481 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6541 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6601 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6661 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6721 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6781 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6841 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     6901 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6961 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7021 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7081 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     7141 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7201 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7261 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7321 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7381 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7441 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7501 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7561 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7621 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7681 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7741 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7801 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7861 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7921 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7981 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8041 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8101 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8161 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8221 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8281 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8341 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8401 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     8461 tcccctatgg tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     8521 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     8581 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agg
//
