LOCUS       V022053                 7730 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022053
VERSION     V022053
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"
     CDS             941..3220
                     /label="TFRC(NM_001128148)"
                     /note="TFRC(NM_001128148)"
                     /gene="TFRC"
     CDS             3227..3292
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3330..3554
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3600..4028
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4042..4371
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4438..5232
                     /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    5406..5527
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5576..5592)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5600..5616
                     /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(5624..5654)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5669..5690
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5978..6563)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6734..7594)
                     /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(7595..7699)
                     /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 aagcttggta ccgagctcgg atccgccacc atgatggatc aagctagatc
      961 agcattctct aacttgtttg gtggagaacc attgtcatat acccggttca gcctggctcg
     1021 gcaagtagat ggcgataaca gtcatgtgga gatgaaactt gctgtagatg aagaagaaaa
     1081 tgctgacaat aacacaaagg ccaatgtcac aaaaccaaaa aggtgtagtg gaagtatctg
     1141 ctatgggact attgctgtga tcgtcttttt cttgattgga tttatgattg gctacttggg
     1201 ctattgtaaa ggggtagaac caaaaactga gtgtgagaga ctggcaggaa ccgagtctcc
     1261 agtgagggag gagccaggag aggacttccc tgcagcacgt cgcttatatt gggatgacct
     1321 gaagagaaag ttgtcggaga aactggacag cacagacttc accggcacca tcaagctgct
     1381 gaatgaaaat tcatatgtcc ctcgtgaggc tggatctcaa aaagatgaaa atcttgcgtt
     1441 gtatgttgaa aatcaatttc gtgaatttaa actcagcaaa gtctggcgtg atcaacattt
     1501 tgttaagatt caggtcaaag acagcgctca aaactcggtg atcatagttg ataagaacgg
     1561 tagacttgtt tacctggtgg agaatcctgg gggttatgtg gcgtatagta aggctgcaac
     1621 agttactggt aaactggtcc atgctaattt tggtactaaa aaagattttg aggatttata
     1681 cactcctgtg aatggatcta tagtgattgt cagagcaggg aaaatcacct ttgcagaaaa
     1741 ggttgcaaat gctgaaagct taaatgcaat tggtgtgttg atatacatgg accagactaa
     1801 atttcccatt gttaacgcag aactttcatt ctttggacat gctcatctgg ggacaggtga
     1861 cccttacaca cctggattcc cttccttcaa tcacactcag tttccaccat ctcggtcatc
     1921 aggattgcct aatatacctg tccagacaat ctccagagct gctgcagaaa agctgtttgg
     1981 gaatatggaa ggagactgtc cctctgactg gaaaacagac tctacatgta ggatggtaac
     2041 ctcagaaagc aagaatgtga agctcactgt gagcaatgtg ctgaaagaga taaaaattct
     2101 taacatcttt ggagttatta aaggctttgt agaaccagat cactatgttg tagttggggc
     2161 ccagagagat gcatggggcc ctggagctgc aaaatccggt gtaggcacag ctctcctatt
     2221 gaaacttgcc cagatgttct cagatatggt cttaaaagat gggtttcagc ccagcagaag
     2281 cattatcttt gccagttgga gtgctggaga ctttggatcg gttggtgcca ctgaatggct
     2341 agagggatac ctttcgtccc tgcatttaaa ggctttcact tatattaatc tggataaagc
     2401 ggttcttggt accagcaact tcaaggtttc tgccagccca ctgttgtata cgcttattga
     2461 gaaaacaatg caaaatgtga agcatccggt tactgggcaa tttctatatc aggacagcaa
     2521 ctgggccagc aaagttgaga aactcacttt agacaatgct gctttccctt tccttgcata
     2581 ttctggaatc ccagcagttt ctttctgttt ttgcgaggac acagattatc cttatttggg
     2641 taccaccatg gacacctata aggaactgat tgagaggatt cctgagttga acaaagtggc
     2701 acgagcagct gcagaggtcg ctggtcagtt cgtgattaaa ctaacccatg atgttgaatt
     2761 gaacctggac tatgagaggt acaacagcca actgctttca tttgtgaggg atctgaacca
     2821 atacagagca gacataaagg aaatgggcct gagtttacag tggctgtatt ctgctcgtgg
     2881 agacttcttc cgtgctactt ccagactaac aacagatttc gggaatgctg agaaaacaga
     2941 cagatttgtc atgaagaaac tcaatgatcg tgtcatgaga gtggagtatc acttcctctc
     3001 tccctacgta tctccaaaag agtctccttt ccgacatgtc ttctggggct ccggctctca
     3061 cacgctgcca gctttactgg agaacttgaa actgcgtaaa caaaataacg gtgcttttaa
     3121 tgaaacgctg ttcagaaacc agttggctct agctacttgg actattcagg gagctgcaaa
     3181 tgccctctct ggtgacgttt gggacattga caatgagttt ctcgaggact acaaagacca
     3241 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtgagg
     3301 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3361 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3421 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3481 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3541 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3601 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3661 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3721 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3781 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3841 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3901 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3961 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4021 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4081 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4141 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4201 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4261 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4321 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4381 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4441 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4501 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4561 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4621 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4681 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4741 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4801 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4861 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4921 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4981 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5041 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5101 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5161 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5221 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5281 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5341 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5401 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5461 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5521 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5581 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5641 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5701 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5761 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5821 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5881 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5941 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6001 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6061 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6121 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6181 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6241 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6301 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6361 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6421 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6481 cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta
     6541 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6601 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6661 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6721 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6781 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6841 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6901 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6961 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7021 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7081 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7141 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7201 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7261 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7321 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7381 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7441 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7501 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7561 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7621 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7681 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
