LOCUS       V032716                 6826 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032716
VERSION     V032716
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             937..2577
                     /label="ELP3(NM_018091)"
                     /note="ELP3(NM_018091)"
                     /gene="ELP3"
     CDS             2590..2655
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2690..2914
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2960..3388
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3402..3731
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3743..4342
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    4502..4623
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4672..4688)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4696..4712
                     /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(4720..4750)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4765..4786
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5074..5659)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5830..6690)
                     /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(6691..6795)
                     /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 aagctttcta gagaattcga tatcggatcc gccaccatga ggcagaagcg gaaaggagat
      961 ctcagccctg ctgagctgat gatgctgact ataggagatg ttattaaaca actgattgaa
     1021 gcccacgagc aggggaaaga catcgatcta aataaggtga aaaccaagac agctgccaaa
     1081 tatggccttt ctgcccagcc ccgcctggtg gatatcattg ctgccgtccc tcctcagtat
     1141 cgcaaggtct tgatgcccaa gttaaaggcg aaacccatca gaactgctag tgggattgct
     1201 gtcgtggctg tgatgtgcaa accccacaga tgtccacaca tcagttttac aggaaatata
     1261 tgtgtatact gccctggtgg acctgattct gattttgagt attccaccca gtcttacact
     1321 ggctatgagc caacctccat gagagctatc cgtgccagat atgacccttt cctacagaca
     1381 agacaccgaa tagaacagtt aaaacaactt ggtcatagtg tggataaagt ggagtttatt
     1441 gtgatgggtg gaacgtttat ggcccttcca gaagaataca gagattattt tattcgaaat
     1501 ttacatgatg ccttatcagg acatacttcc aacaatattt acgaggcagt caagtattct
     1561 gagagaagcc tcacaaagtg tattggaatt actattgaaa ccagaccaga ttactgcatg
     1621 aagcgacatt taagtgacat gttgacctat ggctgcacaa ggctggagat tggggtgcag
     1681 agtgtttatg aagatgtggc tagagacacc aacaggggcc acactgtgaa ggcagtgtgt
     1741 gagtcatttc acctggccaa agattccggt tttaaagtgg tggcccatat gatgcctgac
     1801 ctgccaaacg tgggactaga aagagacatt gaacagttca cagagttttt tgagaaccct
     1861 gcttttcgtc ccgatgggct gaaactctat cctaccctgg tgattcgtgg gaccgggctt
     1921 tatgagcttt ggaaatcagg aagatataag agttactctc ctagtgacct ggttgaattg
     1981 gtggctcgga tcctagccct cgtgcctcca tggactcgag tgtaccgagt acagagggat
     2041 attccaatgc ctttagttag ctcaggagta gagcatggta acctgagaga gctggcactt
     2101 gcaagaatga aagacctcgg aatacagtgt cgagatgtga gaaccagaga agttggaatc
     2161 caagaaattc atcacaaagt acggccatac caggttgaat tggtaaggag agattatgtt
     2221 gcaaatggtg gctgggaaac attcttgtca tacgaagacc cagatcaaga cattttgatt
     2281 ggcctcctac gattacgcaa gtgttcagaa gaaactttcc gtttcgaatt gggtggaggt
     2341 gtctccatag tacgagagct gcatgtgtat gggagtgtgg tccctgtgag cagccgggat
     2401 cctactaaat ttcagcatca gggatttggc atgctgctga tggaggaagc agaaagaata
     2461 gctagagaag aacatgggtc tgggaaaatc gctgtgatat caggggtcgg caccaggaat
     2521 tattatagaa agatcggcta cagattacaa ggcccgtaca tggtgaagat gctgaaactc
     2581 gagaccggtg actacaaaga ccatgacggt gattataaag atcatgacat cgattacaag
     2641 gatgacgatg acaagtgacc ggtaagttta aaccgctgat cagcctcgac tgtgccttct
     2701 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2761 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     2821 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     2881 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     2941 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3001 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3061 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3121 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3181 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3241 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3301 gtctattctt ttgatttata agggattttg gccatttcgg cctattggtt aaaaaatgag
     3361 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3421 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3481 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3541 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3601 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3661 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3721 gcttttgcaa aaagctcccg ggatgaccga gtacaagccc acggtgcgcc tcgccacccg
     3781 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     3841 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     3901 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     3961 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     4021 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     4081 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     4141 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     4201 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     4261 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     4321 gacccgcaag cccggtgcct gattcgaaat gaccgaccaa gcgacgccca acctgccatc
     4381 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4441 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4501 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4561 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4621 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4681 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4741 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4801 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4861 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4921 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4981 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5041 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5101 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5161 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5221 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5281 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5341 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5401 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5461 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5521 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5581 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     5641 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5701 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5761 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5821 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5881 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5941 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6001 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6061 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6121 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6181 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6241 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6301 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6361 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6421 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6481 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6541 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6601 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6661 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6721 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6781 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
