LOCUS       V019571                 7013 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019571
VERSION     V019571
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             671..1693
                     /label="HNRNPA2B1(NM_002137)"
                     /note="HNRNPA2B1(NM_002137)"
                     /gene="HNRNPA2B1"
     CDS             1700..1765
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1789..2377
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2443..2491
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2693..3121
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3135..3464
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3531..4325
                     /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    4501..4622
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4671..4687)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4695..4711
                     /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(4719..4749)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4764..4785
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5073..5661)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5832..6692)
                     /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(6693..6797)
                     /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 taccgccacc atggagagag aaaaggaaca gttccgtaag ctctttattg gtggcttaag
      721 ctttgaaacc acagaagaaa gtttgaggaa ctactacgaa caatggggaa agcttacaga
      781 ctgtgtggta atgagggatc ctgcaagcaa aagatcaaga ggatttggtt ttgtaacttt
      841 ttcatccatg gctgaggttg atgctgccat ggctgcaaga cctcattcaa ttgatgggag
      901 agtagttgag ccaaaacgtg ctgtagcaag agaggaatct ggaaaaccag gggctcatgt
      961 aactgtgaag aagctgtttg ttggcggaat taaagaagat actgaggaac atcaccttag
     1021 agattacttt gaggaatatg gaaaaattga taccattgag ataattactg ataggcagtc
     1081 tggaaagaaa agaggctttg gctttgttac ttttgatgac catgatcctg tggataaaat
     1141 cgtattgcag aaataccata ccatcaatgg tcataatgca gaagtaagaa aggctttgtc
     1201 tagacaagaa atgcaggaag ttcagagttc taggagtgga agaggaggca actttggctt
     1261 tggggattca cgtggtggcg gtggaaattt cggaccagga ccaggaagta actttagagg
     1321 aggatctgat ggatatggca gtggacgtgg atttggggat ggctataatg ggtatggagg
     1381 aggacctgga ggtggcaatt ttggaggtag ccccggttat ggaggaggaa gaggaggata
     1441 tggtggtgga ggacctggat atggcaacca gggtgggggc tacggaggtg gttatgacaa
     1501 ctatggagga ggaaattatg gaagtggaaa ttacaatgat tttggaaatt ataaccagca
     1561 accttctaac tacggtccaa tgaagagtgg aaactttggt ggtagcagga acatgggggg
     1621 accatatggt ggaggaaact atggtccagg aggcagtgga ggaagtgggg gttatggtgg
     1681 gaggagccga tacctcgagg actacaaaga ccatgacggt gattataaag atcatgacat
     1741 cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa tcaacctctg
     1801 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     1861 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     1921 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     1981 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     2041 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     2101 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     2161 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     2221 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     2281 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     2341 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg aggctaactg
     2401 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     2461 ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     2521 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     2581 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     2641 gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc
     2701 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2761 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2821 ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta
     2881 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2941 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3001 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3061 ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3121 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3181 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3241 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3301 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3361 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3421 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3481 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3541 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3601 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3661 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3721 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3781 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3841 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3901 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3961 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4021 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4081 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4141 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4201 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4261 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4321 tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca
     4381 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4441 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4501 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4561 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4621 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4681 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4741 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4801 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4861 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4921 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4981 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5041 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5101 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5161 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5221 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     5281 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5341 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5401 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5461 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     5521 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5581 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5641 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5701 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5761 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5821 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5881 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5941 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6001 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6061 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6121 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6181 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6241 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6301 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6361 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6421 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6481 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6541 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6601 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6661 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6721 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6781 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     6841 gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
     6901 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     6961 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agg
//
