LOCUS       V019791                 7739 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019791
VERSION     V019791
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..2419
                     /label="NCOA5(NM_020967)"
                     /note="NCOA5(NM_020967)"
                     /gene="NCOA5"
     CDS             2426..2491
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2515..3103
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3169..3217
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3419..3847
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3861..4190
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4257..5051
                     /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    5227..5348
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5397..5413)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5421..5437
                     /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(5445..5475)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5490..5511
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5799..6387)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6558..7418)
                     /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(7419..7523)
                     /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 ccatgaatac ggctccatca agacccagcc ccacacgaag
      721 ggatccatat ggctttggag acagtcgaga ttcaaggcgt gatcgatccc caattcgagg
      781 aagtccaagg agagagccca gggatggcag aaatggccgg gatgcccggg acagcagaga
      841 cattcgagac ccccgagact tgcgggacca cagacatagt agagatttgc gggatcacag
      901 agacagcagg agtgtgcgcg acgttcggga cgtgagggat cttagagact ttcgtgatct
      961 aagagactct agggattttc gagatcagcg agaccccatg tacgacagat acagagacat
     1021 gagagactcc cgagatccta tgtacaggag agaaggctct tatgaccgat acctacgaat
     1081 ggatgactat tgcaggagaa aggatgactc ttattttgac cgttacagag atagctttga
     1141 tggacggggc cctccaggcc cagaaagtca gtctcgtgca aaagagcgtt tgaaacgtga
     1201 ggaacggcgt agagaagagc tttatcgtca atattttgag gaaatccaga gacgctttga
     1261 tgccgaaagg cccgttgatt gttctgtgat tgtggtcaac aaacagacaa aagactatgc
     1321 tgagtctgtg gggcggaagg tgcgagacct gggcatggta gtggacttga tcttccttaa
     1381 cacagaagtg tcactgtcac aagccttgga ggatgttagc aggggaggtt ctccttttgc
     1441 tattgtcatc acccagcaac accagattca ccgctcctgc acagtcaaca tcatgtttgg
     1501 aaccccgcaa gagcatcgca acatgcccca agcagatgcc atggtgctgg tggccagaaa
     1561 ttatgagcgt tacaagaatg agtgccggga gaaggaacgt gaggagattg ccagacaggc
     1621 agccaagatg gccgatgaag ccatcctgca ggaaagagag agaggaggcc ctgaggaggg
     1681 agtgcgtggg ggccaccctc cagccatcca gagcctcatc aacctgctgg cagacaacag
     1741 gtacctcact gctgaagaga ctgacaagat catcaactac ctgcgagagc ggaaggagcg
     1801 gctgatgagg agcagcaccg actctctgcc tggcccgatt tcccgccaac cactcggggc
     1861 gacctcgggt gcctcgctga agacacagcc aagctcccaa ccgctccaga gcggccaagt
     1921 gctcccctct gctacaccca ctccatctgc accccccacc tcccagcaag agcttcaggc
     1981 caaaatcctc agcctcttca atagtggcac agtgacggcc aatagcagct ctgcatcccc
     2041 ctcggttgct gccggaaaca ccccaaacca gaatttttcc acagcagcaa acagccagcc
     2101 tcaacaaaga tcacaggctt ctggcaatca gcctccaagc attttgggac agggaggatc
     2161 tgctcagaac atgggcccca gacctggggc tccttcccaa gggctttttg gccagccttc
     2221 cagtcgcctg gcacctgcta gcaacatgac tagccagagg cctgtgtctt ccacaggtat
     2281 caactttgac aatccaagtg tacagaaggc tctggatacc ctgatccaga gtggccctgc
     2341 tctctcccac ctggttagcc agaccacagc acagatgggg cagccacagg cccccatggg
     2401 atcttaccag aggcattacc tcgaggacta caaagaccat gacggtgatt ataaagatca
     2461 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct cgacaatcaa
     2521 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     2581 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     2641 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     2701 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     2761 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     2821 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2881 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     2941 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3001 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     3061 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc
     3121 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     3181 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     3241 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     3301 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     3361 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     3421 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3481 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3541 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     3601 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3661 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3721 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3781 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3841 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3901 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3961 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4021 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4081 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     4141 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     4201 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     4261 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4321 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4381 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     4441 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4501 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4561 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4621 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4681 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4741 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     4801 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4861 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4921 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4981 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5041 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     5101 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5161 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5221 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5281 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5341 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     5401 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5461 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5521 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5581 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5641 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5701 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5761 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5821 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5881 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5941 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     6001 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6061 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6121 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6181 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     6241 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6301 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6361 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6421 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6481 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6541 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6601 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6661 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6721 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6781 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6841 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6901 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6961 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7021 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7081 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7141 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7201 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7261 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7321 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7381 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7441 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7501 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     7561 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat
     7621 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     7681 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
