LOCUS       V035690                 8223 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035690
VERSION     V035690
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..2903
                     /label="NFATC1(NM_172390)"
                     /note="NFATC1(NM_172390)"
                     /gene="NFATC1"
     CDS             2910..2975
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2999..3587
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3653..3701
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3903..4331
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4345..4674
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4741..5535
                     /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    5711..5832
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5881..5897)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5905..5921
                     /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(5929..5959)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5974..5995
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6283..6871)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7042..7902)
                     /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(7903..8007)
                     /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 ccaccatgcc aagcaccagc tttccagtcc
      781 cttccaagtt tccacttggc cctgcggctg cggtcttcgg gagaggagaa actttggggc
      841 ccgcgccgcg cgccggcggc accatgaagt cagcggagga agaacactat ggctatgcat
      901 cctccaacgt cagccccgcc ctgccgctcc ccacggcgca ctccaccctg ccggccccgt
      961 gccacaacct tcagacctcc acaccgggca tcatcccgcc ggcggatcac ccctcggggt
     1021 acggagcagc tttggacggt gggcccgcgg gctacttcct ctcctccggc cacaccaggc
     1081 ctgatggggc ccctgccctg gagagtcctc gcatcgagat aacctcgtgc ttgggcctgt
     1141 accacaacaa taaccagttt ttccacgatg tggaggtgga agacgtcctc cctagctcca
     1201 aacggtcccc ctccacggcc acgctgagtc tgcccagcct ggaggcctac agagacccct
     1261 cgtgcctgag cccggccagc agcctgtcct cccggagctg caactcagag gcctcctcct
     1321 acgagtccaa ctactcgtac ccgtacgcgt ccccccagac gtcgccatgg cagtctccct
     1381 gcgtgtctcc caagaccacg gaccccgagg agggctttcc ccgcgggctg ggggcctgca
     1441 cactgctggg ttccccgcgg cactccccct ccacctcgcc ccgcgccagc gtcactgagg
     1501 agagctggct gggtgcccgc tcctccagac ccgcgtcccc ttgcaacaag aggaagtaca
     1561 gcctcaacgg ccggcagccg ccctactcac cccaccactc gcccacgccg tccccgcacg
     1621 gctccccgcg ggtcagcgtg accgacgact cgtggttggg caacaccacc cagtacacca
     1681 gctcggccat cgtggccgcc atcaacgcgc tgaccaccga cagcagcctg gacctgggag
     1741 atggcgtccc tgtcaagtcc cgcaagacca ccctggagca gccgccctca gtggcgctca
     1801 aggtggagcc cgtcggggag gacctgggca gccccccgcc cccggccgac ttcgcgcccg
     1861 aagactactc ctctttccag cacatcagga agggcggctt ctgcgaccag tacctggcgg
     1921 tgccgcagca cccctaccag tgggcgaagc ccaagcccct gtcccctacg tcctacatga
     1981 gcccgaccct gcccgccctg gactggcagc tgccgtccca ctcaggcccg tatgagcttc
     2041 ggattgaggt gcagcccaag tcccaccacc gagcccacta cgagacggag ggcagccggg
     2101 gggccgtgaa ggcgtcggcc ggaggacacc ccatcgtgca gctgcatggc tacttggaga
     2161 atgagccgct gatgctgcag cttttcattg ggacggcgga cgaccgcctg ctgcgcccgc
     2221 acgccttcta ccaggtgcac cgcatcacag ggaagaccgt gtccaccacc agccacgagg
     2281 ccatcctctc caacaccaaa gtcctggaga tcccactcct gccggagaac agcatgcgag
     2341 ccgtcattga ctgtgccgga atcctgaaac tcagaaactc cgacattgaa cttcggaaag
     2401 gagagacgga catcgggagg aagaacacac gggtacggct ggtgttccgc gttcacgtcc
     2461 cgcaacccag cggccgcacg ctgtccctgc aggtggcctc caaccccatc gaatgctccc
     2521 agcgctcagc tcaggagctg cctctggtgg agaagcagag cacggacagc tatccggtcg
     2581 tgggcgggaa gaagatggtc ctgtctggcc acaacttcct gcaggactcc aaggtcattt
     2641 tcgtggagaa agccccagat ggccaccatg tctgggagat ggaagcgaaa actgaccggg
     2701 acctgtgcaa gccgaattct ctggtggttg agatcccgcc atttcggaat cagaggataa
     2761 ccagccccgt tcacgtcagt ttctacgtct gcaacgggaa gagaaagcga agccagtacc
     2821 agcgtttcac ctaccttccc gccaacggta acgccatctt tctaaccgta agccgtgaac
     2881 atgagcgcgt ggggtgcttt ttcggatccg actacaaaga ccatgacggt gattataaag
     2941 atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa
     3001 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     3061 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     3121 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     3181 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     3241 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     3301 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     3361 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     3421 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     3481 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     3541 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg
     3601 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     3661 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     3721 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     3781 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     3841 cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc
     3901 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3961 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4021 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     4081 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4141 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4201 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4261 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4321 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4381 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4441 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4501 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4561 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     4621 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4681 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     4741 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4801 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4861 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4921 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4981 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5041 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5101 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5161 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5221 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5281 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5341 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5401 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5461 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5521 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     5581 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5641 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5701 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5761 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5821 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5881 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5941 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6001 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6061 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6121 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6181 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6241 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6301 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6361 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6421 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     6481 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6541 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6601 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6661 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6721 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6781 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6841 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6901 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6961 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7021 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7081 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7141 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7201 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7261 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7321 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7381 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7441 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7501 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7561 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7621 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7681 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7741 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7801 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7861 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7921 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7981 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     8041 cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc
     8101 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     8161 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     8221 agg
//
