LOCUS       V041224                 6747 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041224
VERSION     V041224
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        61..364
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        365..568
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             641..2599
                     /label="PIAS1(NM_001320687)"
                     /note="PIAS1(NM_001320687)"
                     /gene="PIAS1"
     CDS             2600..2617
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     CDS             2618..2671
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             2693..3412
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     polyA_signal    3535..3656
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(3663..4118)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4145..4249
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        4251..4608
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4643..5437
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5669..5716
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6045..6633
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcgccacc atgtttacac tccaagattc
      661 gtatgttaag caaatggtta tgagccttag agtttctgaa ctccaagtac tgttgggcta
      721 cgccgggaga aacaagcacg gacgcaaaca cgaacttctc acaaaagccc tgcatttgct
      781 aaaggctggc tgtagtcctg ctgtgcaaat gaaaattaag gaactctata ggcggcggtt
      841 cccacagaaa atcatgacgc ctgcagactt gtccatcccc aacgtacatt caagtcctat
      901 gccagcaact ttgtctccat ctaccattcc acaactcact tacgatggtc accctgcatc
      961 atcgccatta ctccctgttt ctcttctggg acctaaacat gaactggaac tcccacatct
     1021 tacatcagct cttcacccag tccatccgga tataaaactt caaaaattac cattttatga
     1081 tttactggat gaactgataa aacccaccag tctagcatca gacaacagtc agcgctttcg
     1141 agaaacctgt tttgcatttg ccttgacacc acaacaagtg cagcaaatca gtagttccat
     1201 ggatatttct gggaccaaat gtgacttcac agtacaggtc cagttaaggt tttgtttatc
     1261 agaaaccagt tgtccacaag aagatcactt cccacccaat ctttgtgtga aagtgaatac
     1321 aaaaccttgc agccttccag gttaccttcc acctacaaaa aatggcgtgg aaccaaagcg
     1381 acccagccga ccaattaata tcacctcact tgtccgactg tccacaacag taccaaacac
     1441 gattgttgtt tcttggactg cagaaattgg aagaaactat tccatggcag tatatcttgt
     1501 aaaacagttg tcctcaacag ttcttcttca gaggttacga gcaaagggaa taaggaatcc
     1561 ggatcattct agagctttaa ttaaagagaa gttgactgcg gatccagaca gtgaaatagc
     1621 tacaaccagc ctaagggttt ctctactatg tccacttggt aaaatgcggc tgacaattcc
     1681 gtgtcgggcc cttacatgtt ctcatctaca atgttttgac gcaactcttt acattcagat
     1741 gaatgagaaa aaaccaacct gggtttgtcc tgtctgtgat aagaaggctc catatgaaca
     1801 ccttattatt gatggcttgt ttatggaaat cctaaagtac tgtacagact gtgatgaaat
     1861 acaatttaag gaggatggca cttgggcacc gatgagatca aaaaaggaag tacaggaagt
     1921 ttctgcctct tacaatggag tcgatggatg cttgagctcc acattggagc atcaggtagc
     1981 gtctcaccac cagtcctcaa ataaaaacaa gaaagtagaa gtgattgacc taaccataga
     2041 cagttcatct gatgaagagg aagaagagcc atctgccaag aggacctgtc cttccctatc
     2101 tcccacatca ccactaaata ataaaggcat tttaagtctt ccacatcaag catctccagt
     2161 atcccgcacc ccaagccttc ctgctgtaga cacaagctac attaatacct ccctcatcca
     2221 agactatagg catcctttcc acatgacacc catgccttac gacttacaag gattagattt
     2281 ctttcctttc ttatcaggag acaatcagca ttacaacacc tccttgcttg ccgctgcagc
     2341 agcagcagtt tcagatgatc aagacctcct acactcgtct cggtttttcc cgtatacctc
     2401 ctcacagatg tttcttgatc agttaagtgc aggaggcagt acttctctgc caaccaccaa
     2461 tggaagcagt agtggcagta acagcagcct ggtttcttcc aacagcctaa gggaaagcca
     2521 tagccacacc gtcacaaaca ggagcagcac ggacacggca tccatctttg gcatcatacc
     2581 agacattatt tcattggacc atcaccatca ccatcacgag ggcagaggaa gtcttctaac
     2641 atgcggtgac gtggaggaga atcccggccc tgcggatcca ccggtcgcca ccatggtgag
     2701 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     2761 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     2821 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     2881 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     2941 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     3001 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     3061 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     3121 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     3181 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     3241 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     3301 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     3361 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg
     3421 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     3481 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3541 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3601 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     3661 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3721 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3781 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3841 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3901 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3961 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     4021 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     4081 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     4141 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4201 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     4261 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4321 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4381 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4441 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4501 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4561 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     4621 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4681 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4741 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4801 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     4861 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4921 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4981 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     5041 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5101 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5161 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5221 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5281 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5341 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5401 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5461 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5521 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5581 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     5641 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     5701 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     5761 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5821 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5881 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5941 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6001 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6061 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6121 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6181 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6241 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6301 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6361 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6421 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6481 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6541 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6601 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6661 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6721 gtggataacc gtattaccgc catgcat
//
