LOCUS       V045488                 8682 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045488
VERSION     V045488
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..3362
                     /label="Hlcs(NM_001360045)"
                     /note="Hlcs(NM_001360045)"
                     /gene="Hlcs"
     CDS             3369..3434
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3458..4046
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4112..4160
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4362..4790
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4804..5133
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5200..5994
                     /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    6170..6291
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6340..6356)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6364..6380
                     /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(6388..6418)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6433..6454
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6742..7330)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7501..8361)
                     /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(8362..8466)
                     /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 ccaccatgct catcacgctg tgctacctgt
      781 acctgtgggc gcgctggggg cgccggccgg ccgcgctggt gctgtccacg gtgcggcggc
      841 tgcgtgcttc gcgctgctcc ttcaccttct gccgcgcggc cgcgcagccc agggacgcgc
      901 gcgtgtgcct gagccagggc ggccgcgtct tctgcgtcgg cgagagccag tctgttgatg
      961 acttgaacaa atgggccctg tttctcgtgt ctcccttcat acttgaggca gaacatatag
     1021 catttgtgac agagagcatt tggatacaag gtaccagttt acaagagcca tcatcttcag
     1081 aaaccgttgt aaaatggtct gactgctgtc tgccgttagc ctgccgaccc ggggacccct
     1141 accagttaat tgctaaggca agcgtggaca acttcagcaa gcttggggtg gccttcatgg
     1201 aggacaggct ccagatggac aacggcctga tagcccagaa gatcgtttca gtccacttaa
     1261 aggaccctgc tctgaaggaa ctcggcaaag cctccgacaa gcaagtgcag ggcccgcccc
     1321 cgggcccaga agcttccccc gaggcccagc ctgcacaagg tgtcatggag catgctggcc
     1381 aaggtgattg caaggccgcc ggtgaaggcc cttccccacg gaggaggggc tgtgcccctg
     1441 aaagtgagcc tgctgcagat ggggacccag ggctgagctc cccggagctc tgtcagctcc
     1501 acctgtccat ctgtcacgag tgtctggagc tcgagaacag taccattgac tcggtgaggt
     1561 ctgcgtctgc ggagaacatt ccagaccttc cctgtgatca cagcggcgtg gagggtgctg
     1621 cgggcgaact ctgccccgaa cggaaaggga agagagtcaa catttcggga aaggcgccca
     1681 atatcttgct gtatgtgggc tctggttccg aggaagccct gggccggctc cagcaggtgc
     1741 gatctgtcct gactgactgt gtggacacgg acagctacac tctctaccac ctgctggaag
     1801 acagcgctct gagggacccg tggtctgaca actgcctcct gctagttatt gccagcaggg
     1861 accccatccc caaagacata cagcacaagt tcatggccta tctttctcag ggagggaagg
     1921 tgctgggcct gtcttccccc ttcacgttgg gtggctttcg ggtgaccagg agagatgtgc
     1981 tacggaacac agtccagaac ttggttttct ctaaggctga tgggactgag gtacggctca
     2041 gtgtcctgag cagcggctac gtttatgaag aaggcccgag cttgggcagg ctccagggcc
     2101 acctggagaa tgaggacaag gataaaatga ttgtgcacgt gccttttgga accctcggag
     2161 gggaagccgt tctctgccag gtgcacctgg agctcccgcc cggcgcctcc ctggtgcaga
     2221 ctgcagacga ctttaacgtg ctcaaatcca gcaatgtgag aagacacgag gtccttaagg
     2281 agatcctgac tgccctcggt ctgagctgtg acgcaccaca ggttcctgcc ttaacgcccc
     2341 tgtacttact gctggccgct gaggaaaccc aggatccgtt catgcagtgg ctcgggaggc
     2401 acacagatcc tgaaggaatc ataaaatcca gcaagctctc ccttcagttt gtttcctcct
     2461 acacatctga agcggagatc accccgtctt ccatgcctgt ggtcactgac ccagaagcct
     2521 tttcatcaga gcacttcagc ctagagacct atcgacagaa tctgcagacc acgcgccttg
     2581 ggaaagtgat tctgtttgca gaagtgacct ccaccaccat gagtctcctg gatgggttga
     2641 tgtttgagat gccgcaggaa atgggcttaa ttgccattgc agttcggcag acccagggca
     2701 aaggaagggg cccgaatgcc tggctgagcc ccgtgggatg tgctctttcc actctgctgg
     2761 tcttcatccc cctgaggtca caactgggac agaggattcc atttgtccag catttgatgt
     2821 ccttggctgt ggtggaggca gtgcggtcta tcccagggta tgaggatatc aacttgcgag
     2881 tgaagtggcc caacgatatt tattacagcg atctcatgaa gatcggtggg gttctggtta
     2941 attcaacact tatgggagaa acattttata tcctcatcgg ctgtggattc aatgtgacga
     3001 acagtaatcc taccatttgt atcaacgacc tcatcgaaga acacaataag cagcacgggg
     3061 cgggcctgaa gcccctgcga gcggactgtc tcatagccag agccgtgacc gtgctggaga
     3121 aactgatcga caggtttcag gaccaagggc ccgacggcgt tcttcccctc tattataaat
     3181 actgggtgca cggtggtcag caagtccgcc tgggcagcac ggaggggcca caggcatcta
     3241 ttgtgggcct tgatgactcc gggttcctgc aggtccacca agaggatggt ggggttgtga
     3301 cggtgcatcc agatggcaac tccttcgaca tgctgagaaa cctcatcgtg cccaagcgac
     3361 agggatccga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     3421 atgacgatga caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat
     3481 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     3541 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     3601 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     3661 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3721 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3781 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3841 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     3901 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3961 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     4021 gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag
     4081 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     4141 gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     4201 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     4261 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     4321 catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     4381 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4441 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4501 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     4561 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4621 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4681 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     4741 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4801 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4861 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4921 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4981 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5041 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     5101 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     5161 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     5221 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     5281 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     5341 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     5401 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     5461 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     5521 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5581 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5641 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5701 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     5761 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5821 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5881 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5941 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     6001 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     6061 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     6121 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     6181 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6241 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     6301 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6361 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6421 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6481 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6541 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6601 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6661 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6721 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6781 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6841 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6901 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     6961 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7021 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7081 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7141 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     7201 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7261 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7321 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7381 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7441 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7501 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7561 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7621 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7681 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7741 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7801 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7861 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7921 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7981 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8041 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8101 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8161 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8221 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8281 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8341 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8401 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8461 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     8521 cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     8581 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     8641 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg
//
