LOCUS       V044356                 9543 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044356
VERSION     V044356
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..4223
                     /label="CILP2(NM_153221)"
                     /note="CILP2(NM_153221)"
                     /gene="CILP2"
     CDS             4230..4295
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4319..4907
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4973..5021
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5223..5651
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5665..5994
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6061..6855
                     /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    7031..7152
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7201..7217)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7225..7241
                     /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(7249..7279)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7294..7315
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7603..8191)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8362..9222)
                     /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(9223..9327)
                     /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 ccaccatggc gtcgctgctg ccactgctct
      781 gtctctgtgt cgtcgctgcg cacctggcgg gggcccgaga cgccaccccc accgaggagc
      841 caatggcgac tgcactgggc ctggaaagac ggtccgtgta caccggccag ccctcaccag
      901 ccctggagga ctgggaagag gccagcgagt ggacgtcctg gttcaacgtg gaccaccccg
      961 gaggcgacgg cgacttcgag agcctggctg ccatccgctt ctactacggg ccagcgcgcg
     1021 tgtgcccgcg accgctggcg ctggaagcgc gcaccacgga ctgggccctg ccgtccgccg
     1081 tcggcgagcg cgtgcacttg aaccccacgc gcggcttctg gtgcctcaac cgcgagcaac
     1141 cgcgtggccg ccgctgctcc aactaccacg tgcgcttccg ctgcccacta gaagcctcgt
     1201 ggggcgcgtg gggcccgtgg ggtccctgct cggggagctg tgggccaggc cgtcgcttgc
     1261 gccgccgcca ctgcccaagc cccgctgggg atgcgtgtcc cgggcgtcct ctggaggcgc
     1321 agaagtgcgt gcggcctcgg tgtccagggt gcagccttga cacctgtgaa tgcccggacc
     1381 acatcctcct gggctcggtg gtcaccccat ctgggcaacc actgctagga gccagggtct
     1441 ccctgcgaga ccagcctggc actgtggcca ccagcgatgc tcacggaacc ttccgggtgc
     1501 ctggtgtctg tgctgacagc cgcgccaaca tcagggccca gatggatggc ttctctgcag
     1561 gggaggccca ggcccaggcc aacggatcca tctctgtggt caccatcatc cttgataagt
     1621 tggagaagcc gtacctggtg aaacaccctg agtcccgagt gcgagaggct ggccagaatg
     1681 tgactttctg ctgcaaagcc tccgggaccc ccatgcccaa gaaatactcc tggttccaca
     1741 atgggaccct gctggacagg cgagctcatg ggtacggggc ccacctggag ctgcggggac
     1801 tgcgcccaga ccaggctggc atctaccact gcaaggcatg gaatgaggcg ggtgccgtgc
     1861 gctcgggcac tgcccggctc actgtacttg ccccaggcca gccagcctgc gacccccggc
     1921 cccgagagta cctgatcaag ctccctgagg actgtggtca gccaggtagt ggccctgcct
     1981 acctggatgt gggcctctgt cccgacaccc gctgccccag cctggcaggc tccagccccc
     2041 gctgcgggga cgccagctcc cgctgctgct ctgtgcgccg tctggagaga agggagattc
     2101 actgccctgg ctacgtcctc ccagtgaagg tggtggcaga gtgtggctgc cagaagtgtc
     2161 tgccccctcg ggggctggtc cggggccgtg ttgtggctgc tgactccggg gagccgctac
     2221 gcttcgccag gattctgctg ggccaggagc ccatcggctt caccgcctac cagggcgact
     2281 ttaccattga ggtgccgccc tccacccagc ggctggtggt gacttttgtg gaccccagcg
     2341 gtgagttcat ggacgctgtc cgggtcttgc cttttgatcc tcgaggtgcc ggcgtgtacc
     2401 acgaggtcaa ggccatgcgg aagaaagccc cggtcatttt acataccagc cagagcaaca
     2461 cgatccccct gggcgagctg gaagatgagg cgcccctggg cgagctggtc ctgccttctg
     2521 gcgctttccg cagagccgac ggcaaaccct actcggggcc tgtggaggcc cgggtgacgt
     2581 tcgtggaccc ccgagacctc acctcggcgg cgtctgcccc cagtgacctg cgcttcgtgg
     2641 acagcgacgg cgagctggct ccactgcgca cctacggcat gttctccgtg gacctccgtg
     2701 cgcccggctc cgcggagcag ctgcaggtgg ggccggtggc cgtgcgggtg gccgccagcc
     2761 agatccacat gccaggccac gtggaggccc tcaagctgtg gtcgctgaac cccgagaccg
     2821 gcttgtggga ggaggagagc ggcttccggc gcgaggggtc ctcgggcccc cgggtgcgcc
     2881 gggaggagcg cgtcttcctg gtgggcaacg tggagatccg ggagcggcgc ctgttcaatc
     2941 tggacgtgcc tgagcgccgc cgctgcttcg tgaaggtgcg cgcctacgcc aacgacaagt
     3001 tcacccccag cgagcaggtg gagggcgtgg tggtcacgct ggtcaatctg gagcccgccc
     3061 ccggcttctc cgccaacccc cgtgcctggg gccgctttga cagcgcggtc accggcccca
     3121 atggcgcctg cctccccgcc ttctgcgacg ccgacaggcc agacgcctac accgccctgg
     3181 tcaccgccac cctgggcggc gaggagctgg agccggcccc ttccttgccc cgcccactcc
     3241 cggccaccgt gggcgtcacc cagccctacc tggacaggct ggggtaccgt cggacggacc
     3301 acgacgatcc cgccttcaag cgtaacggct tccgcatcaa cctcgccaag cccaggccag
     3361 gtgaccccgc cgaggccaat gggcctgtgt acccgtggcg cagcctgcgg gaatgccagg
     3421 gggccccggt gactgccagc cacttccgct tcgccagggt ggaggcggac aagtacgagt
     3481 acaacgtggt ccccttccga gagggcacac ctgcctcctg gactggcgat ctcctggcct
     3541 ggtggcccaa cccgcaggag ttccgggcct gcttcctcaa ggtgaagatc cagggtcccc
     3601 aggagtatat ggtccgctcc cacaacgcag ggggcagcca cccacgcacc cgcggccagc
     3661 tctacggact tcgggatgcc cggagtgtgc gagaccccga gcgtccgggc acctcggcag
     3721 cctgcgtgga gttcaagtgc agcgggatgc tgttcgacca gcggcaggtg gacaggacgc
     3781 tggtgaccat tatgccccag ggcagctgcc ggcgcgtggc cgtcaacgga ctccttcggg
     3841 attacctgac ccggcacccc ccaccggtgc ccgcggagga cccagctgcc ttctccatgc
     3901 tggcccccct agaccctctg ggccacaact atggcgtcta cactgtcact gaccagagcc
     3961 cacgcttggc caaggagatc gccattggcc gctgctttga tggttcctct gacggcttct
     4021 ccagagagat gaaggctgat gccggcacag ccgtcacctt ccagtgccgg gagccaccgg
     4081 ccggacgacc cagcctcttc cagaggctgc tggagtcccc ggcgacagca cttggtgaca
     4141 tccgcaggga gatgagcgag gcggcgcagg cacaggcccg ggcctcaggt cccctccgca
     4201 cccgccgggg tagggtccgg cagggatccg actacaaaga ccatgacggt gattataaag
     4261 atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa
     4321 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     4381 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     4441 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     4501 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     4561 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     4621 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     4681 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     4741 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     4801 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     4861 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg
     4921 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     4981 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     5041 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     5101 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     5161 cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc
     5221 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5281 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5341 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     5401 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5461 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5521 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5581 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5641 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     5701 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     5761 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5821 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5881 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     5941 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     6001 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     6061 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     6121 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     6181 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     6241 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     6301 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     6361 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     6421 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     6481 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     6541 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     6601 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6661 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6721 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     6781 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6841 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     6901 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     6961 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     7021 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     7081 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     7141 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     7201 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     7261 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     7321 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     7381 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     7441 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     7501 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7561 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7621 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7681 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7741 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     7801 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7861 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7921 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7981 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     8041 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     8101 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     8161 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     8221 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     8281 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     8341 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     8401 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     8461 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     8521 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8581 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8641 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8701 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8761 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8821 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8881 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8941 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     9001 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     9061 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     9121 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     9181 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     9241 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     9301 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     9361 cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc
     9421 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     9481 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     9541 agg
//
