LOCUS       V044307                 7465 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044307
VERSION     V044307
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             706..2124
                     /label="CAMK4(NM_001744)"
                     /note="CAMK4(NM_001744)"
                     /gene="CAMK4"
     CDS             2131..2157
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2161..2187
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2191..2217
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2241..2829
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2895..2943
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3145..3573
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3587..3916
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3983..4777
                     /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    4953..5074
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5123..5139)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5147..5163
                     /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(5171..5201)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5216..5237
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5525..6113)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6284..7144)
                     /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(7145..7249)
                     /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 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgct caaagtcacg
      721 gtgccctcct gctccgcctc gtcctgctct tcggtcaccg ccagtgcggc cccggggacc
      781 gcgagcctcg tcccggatta ctggatcgac ggctccaaca gggatgcgct gagcgatttc
      841 ttcgaggtgg agtcggagct gggacggggt gctacatcca ttgtgtacag atgcaaacag
      901 aaggggaccc agaagcctta tgctctcaaa gtgttaaaga aaacagtgga caaaaaaatc
      961 gtaagaactg agataggagt tcttcttcgc ctctcacatc caaacattat aaaacttaaa
     1021 gagatatttg aaacccctac agaaatcagt ctggtcctag aactcgtcac aggaggagaa
     1081 ctgtttgata ggattgtgga aaagggatat tacagtgagc gagatgctgc agatgccgtt
     1141 aaacaaatcc tggaggcagt tgcttatcta catgaaaatg ggattgtcca tcgtgatctc
     1201 aaaccagaga atcttcttta tgcaactcca gccccagatg caccactcaa aatcgctgat
     1261 tttggactct ctaaaattgt ggaacatcaa gtgctcatga agacagtatg tggaacccca
     1321 gggtactgcg cacctgaaat tcttagaggt tgtgcctatg gacctgaggt ggacatgtgg
     1381 tctgtaggaa taatcaccta catcttactt tgtggatttg aaccattcta tgatgaaaga
     1441 ggcgatcagt tcatgttcag gagaattctg aattgtgaat attactttat ctccccctgg
     1501 tgggatgaag tatctctaaa tgccaaggac ttggtcagaa aattaattgt tttggatcca
     1561 aagaaacggc tgactacatt tcaagctctc cagcatccgt gggtcacagg taaagcagcc
     1621 aattttgtac acatggatac cgctcaaaag aagctccaag aattcaatgc ccggcgtaag
     1681 cttaaggcag cggtgaaggc tgtggtggcc tcttcgcgcc tgggaagtgc cagcagcagc
     1741 catggcagca tccaggagag ccacaaggct agccgagacc cttctccaat ccaagatggc
     1801 aacgaggaca tgaaagctat tccagaagga gagaaaattc aaggcgatgg ggcccaagcc
     1861 gcagttaagg gggcacaggc tgagctgatg aaggtgcaag ccttagagaa agttaaaggt
     1921 gcagatataa atgctgaaga ggcccccaaa atggtgccca aggcagtgga ggatgggata
     1981 aaggtggctg acctggaact agaggagggc ctagcagagg agaagctgaa gactgtggag
     2041 gaggcagcag ctcccagaga agggcaagga agctctgctg tgggttttga agttccacag
     2101 caagatgtga tcctgccaga gtacctcgag tacccatacg acgtacctga ttatgcaact
     2161 tatccttatg atgtacctga ttatgctagc tatccttatg atgtcccaga ctacgcatag
     2221 tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     2281 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     2341 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     2401 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     2461 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     2521 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     2581 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
     2641 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     2701 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     2761 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     2821 cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     2881 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg
     2941 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     3001 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     3061 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca
     3121 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3181 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3241 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3301 gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3361 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3421 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3481 tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa
     3541 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3601 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3661 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3721 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3781 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3841 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3901 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     3961 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4021 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4081 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4141 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4201 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4261 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4321 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4381 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4441 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4501 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4561 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4621 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4681 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4741 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa
     4801 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4861 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4921 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     4981 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5041 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5101 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5161 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5221 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5281 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5341 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5401 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5461 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5521 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5581 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5641 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5701 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     5761 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5821 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5881 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5941 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6001 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6061 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6121 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6181 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6241 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6301 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6361 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6421 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6481 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6541 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6601 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6661 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6721 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6781 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6841 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6901 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6961 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7021 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7081 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7141 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7201 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7261 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct
     7321 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     7381 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     7441 caattgcatg aagaatctgc ttagg
//
