LOCUS       V032024                 7209 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032024
VERSION     V032024
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..1889
                     /label="HNRNPA3(NM_194247)"
                     /note="HNRNPA3(NM_194247)"
                     /gene="HNRNPA3"
     CDS             1896..1961
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1985..2573
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2639..2687
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2889..3317
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3331..3660
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3727..4521
                     /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    4697..4818
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4867..4883)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4891..4907
                     /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(4915..4945)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4960..4981
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5269..5857)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6028..6888)
                     /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(6889..6993)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgga ggtaaaaccg ccgcccggtc
      781 gcccccagcc cgactccggc cgtcgccgtc gccgccgggg ggaggagggc catgatccaa
      841 aggaaccaga gcagttgaga aaactgttta ttggtggtct gagctttgaa actacagatg
      901 atagtttacg agaacatttt gagaaatggg gcacactcac agattgtgtg gtaatgagag
      961 acccccaaac aaaacgttcc aggggctttg gttttgtgac ttattcttgt gttgaagagg
     1021 tggatgcagc aatgtgtgct cgaccacaca aggttgatgg gcgtgtagtg gaaccaaaga
     1081 gagctgtttc tagagaggat tctgtaaagc ctggtgccca tctaacagtg aagaaaattt
     1141 ttgttggtgg tattaaagaa gatacagaag aatataattt gagagactac tttgaaaagt
     1201 atggcaagat tgaaaccata gaagttatgg aagacaggca gagtggaaaa aagagaggat
     1261 ttgcttttgt aacttttgat gatcatgata cagttgataa aattgttgtt cagaaatacc
     1321 acactattaa tgggcataat tgtgaagtga aaaaggccct ttctaaacaa gagatgcagt
     1381 ctgctggatc acagagaggt cgtggaggtg gatctggcaa ttttatgggt cgcggaggga
     1441 actttggagg tggtggaggt aattttggcc gtggtggaaa ctttggtgga agaggaggct
     1501 atggtggtgg aggtggtggc agcagaggta gttatggagg aggtgatggt ggatataatg
     1561 gatttggagg tgatggtggc aactatggcg gtggtcctgg ttatagtagt agagggggct
     1621 atggtggtgg tggaccagga tatggaaacc aaggtggtgg atatggtgga ggtggaggat
     1681 atgatggtta caatgaagga ggaaattttg gcggtggtaa ctatggtggt ggtgggaact
     1741 ataatgattt tggaaattat agtggacaac agcaatcaaa ttatggaccc atgaaagggg
     1801 gcagttttgg tggaagaagc tcgggcagtc cctatggtgg tggttatgga tctggtggtg
     1861 gaagtggtgg atatggtagc agaaggttcg gatccgacta caaagaccat gacggtgatt
     1921 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct
     1981 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     2041 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     2101 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     2161 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     2221 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     2281 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     2341 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     2401 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     2461 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     2521 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     2581 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     2641 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     2701 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     2761 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     2821 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     2881 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2941 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3001 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     3061 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3121 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3181 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3241 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     3301 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3361 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3421 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3481 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3541 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3601 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3661 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     3721 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3781 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3841 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3901 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3961 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4021 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4081 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4141 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4201 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4261 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4321 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4381 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4441 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4501 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     4561 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4621 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4681 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4741 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4801 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     4861 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4921 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4981 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5041 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5101 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5161 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5221 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5281 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5341 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5401 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5461 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5521 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5581 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5641 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5701 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5761 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5821 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5881 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5941 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6001 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6061 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6121 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6181 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6241 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6301 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6361 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6421 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6481 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6541 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6601 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6661 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6721 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6781 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6841 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6901 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6961 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7021 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     7081 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     7141 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     7201 ctgcttagg
//
