LOCUS       V025174                 7780 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025174
VERSION     V025174
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..3219
                     /label="LRCH1(NM_001164211)"
                     /note="LRCH1(NM_001164211)"
                     /gene="LRCH1"
     CDS             3226..3252
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3256..3282
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3286..3312
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    3347..3571
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3617..4045
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4059..4388
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4455..5249
                     /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    5423..5544
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5593..5609)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5617..5633
                     /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(5641..5671)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5686..5707
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5995..6580)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6751..7611)
                     /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(7612..7716)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atggcgacgc cgggaagcga accccaacct
      961 ttcgtcccgg ccctttcggt agctactctg cacccacttc atcatcccca ccaccaccac
     1021 caccaccatc agcaccacgg aggaaccggc gcccccggcg gggcgggtgg tggcggcggt
     1081 ggcagcgggg gcttcaacct gcccttgaac cggggtctgg agcgcgcgct tgaggaggcg
     1141 gccaactccg gggggctgaa cctgagcgcc aggaaattga aggaatttcc ccgtaccgca
     1201 gcccccgggc acgacctctc ggacacggtg caggcagact tatctaaaaa cagactggtt
     1261 gaagttccaa tggaattgtg ccattttgta tcactggaaa ttcttaatct gtatcacaac
     1321 tgtatcagag tcattcctga ggccatcgtt aatctgcaga tgctgactta cctgaacttg
     1381 agtcgaaatc agctgtccgc cctgcctgcc tgcctgtgtg gtctgcctct caaagtctta
     1441 atcgcaagta acaacaaact tggatcatta ccagaagaga taggtcagct caaacagtta
     1501 atggagctgg atgtcagctg caacgagatc acagcgttgc cccagcagat aggtcagttg
     1561 aaatctctac gagaactgaa tgtcagaaga aattacctta aagttttacc acaagaacta
     1621 gtagatcttt ccttggtaaa gtttgacttt tcctgcaaca aagtgctcgt gattccaatt
     1681 tgttttagag agatgaagca gctgcaagtg ttactacttg agaataaccc tctgcagtct
     1741 cctccagcac agatttgcac aaagggcaaa gttcacatat ttaagtatct gagcatacaa
     1801 gcatgccaga ttaagacagc tgactccctt tatctccaca ccatggagag gccacattta
     1861 caccagcacg tggaagatgg caagaaggat tctgattcgg gagttggaag tgataatgga
     1921 gataagcgat tatctgccac cgagccttct gacgaagaca ctgttagcct caatgtgcca
     1981 atgtcaaaca tcatggaaga agaacagatc atcaaggagg actcgtgcca tcgccttagc
     2041 cccgttaaag gggaatttca tcaggaattt caaccggagc cttccctttt gggtgacagc
     2101 accaactcag gagaagaaag agaccagttt actgatagag cagatggtct ccattcggaa
     2161 tttatgaact ataaggcaag ggcagaagac tgtgaagagc tgttacggat agaagaggat
     2221 gtgcactggc aaactgaggg cataataagt tcatccaaag atcaggacat ggatatagca
     2281 atgatcgagc agctgagaga agcagtagat ttgctgcaag atcccaatgg attaagcaca
     2341 gatattacag agagaagtgt tttaaaccta tatcctatgg gatcagcaga agccttagaa
     2401 ttacaagatt ctgcactgaa tggtcaaata cagctggaga catctccggt gtgtgaggtg
     2461 caaagtgatc taacattaca gagtaacggg agccagtatt ctccaaatga gattagagag
     2521 aactcccctg cagtctctcc taccacaaac agcacagctc catttggcct gaagcctcga
     2581 tcagacccag ccctcattct tcctcctatc tccttcaaca cacttacaca ggcacagaca
     2641 tgggacagct ctagctacag tgttccatct gaaggagaca gtgacaatgt gtttctaaga
     2701 cctcagagaa atttggaatc tatagacccg cagtttacaa tccggaggaa aatggagcag
     2761 atgagagaag agaaagagct ggtggaacaa cttcgtgaga gcattgagat gagattgaag
     2821 gtcagtctac acgaagacct gggggcagcc ctcatggatg gtgtcgtcct ctgccatctg
     2881 gtcaaccaca tccgcccacg gtcggttgca agcatccatg tcccatcacc agcggttccc
     2941 aaacttagca tggccaaatg cagaagaaat gtggaaaact ttttggaagc atgccgaaaa
     3001 ttaggagtac cagaggctga cctctgctct ccgtgtgaca tcctgcagtt ggattttcgt
     3061 cacattcgaa agactgttga cactctgctg gcactcgggg agaaagcccc accaccaact
     3121 tctgccctcc gctccaggga ccttataggc ttctgtcttg tccatattct ctttatagtg
     3181 ctggtctata tcacttacca ctggaatgct ctgtccgcac tcgagtaccc atacgacgta
     3241 cctgattatg caacttatcc ttatgatgta cctgattatg ctagctatcc ttatgatgtc
     3301 ccagactacg catgagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     3361 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3421 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3481 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3541 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3601 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3661 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3721 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3781 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3841 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3901 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3961 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4021 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4081 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4141 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4201 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4261 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4321 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4381 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4441 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4501 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4561 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4621 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4681 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4741 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4801 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4861 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4921 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4981 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5041 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5101 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5161 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5221 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5281 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5341 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5401 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5461 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5521 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5581 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5641 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5701 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5761 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5821 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5881 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5941 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6001 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6061 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6121 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6181 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6241 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6301 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6361 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6421 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6481 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     6541 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6601 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6661 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6721 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6781 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6841 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6901 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6961 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7021 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7081 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7141 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7201 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7261 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7321 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7381 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7441 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7501 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7561 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7621 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7681 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7741 tgacgtcgac ggatcgggag atctcccgat cccctatggt
//
