LOCUS       V047295                 8265 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047295
VERSION     V047295
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..2945
                     /label="Lrrn2(NM_010732)"
                     /note="Lrrn2(NM_010732)"
                     /gene="Lrrn2"
     CDS             2952..3017
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3041..3629
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3695..3743
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3945..4373
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4387..4716
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4783..5577
                     /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    5753..5874
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5923..5939)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5947..5963
                     /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(5971..6001)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6016..6037
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6325..6913)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7084..7944)
                     /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(7945..8049)
                     /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 ccaccatgag gctcctcgtg gctgccctct
      781 tgctatcttg ggtggctggt accaccgctg cagcaccggt ggtaccctgg cgtgtgccct
      841 gccctcctca gtgtgcctgc cagatccggc cctggtacac accccgctcg tcctaccgtg
      901 aagccaccac tgtcgactgc aatgacctct tcctgacggc agtgccccca aggctccctg
      961 cggggacaca gaccctgctc ctgcagagta acagcatcag ccggatagac cagactgagc
     1021 ttgcctactt ggccaacctc acagagctgg acctgtccca gaacagcttc tcagatgccc
     1081 gagactgcga tttccaagcc ctgcctcagc tgctgagcct acatctagaa gagaaccggc
     1141 tgaaccggct ggaggatcac agcttcgctg ggctgaccag cctgcaggag ctctatctca
     1201 accataacca gctctgccgc atctccccca gggcctttgc agggctgggt aacctgctga
     1261 gactgcacct caattccaac ctgcttagga ccatcgacag ccgctggttc gagatgctgc
     1321 ccaacttgga aatcctcatg attggcggca acaaggtgga tgccatcttg gacatgaact
     1381 tccggcccct ggccaacctg cgcagcctgg tgctggcagg catgagcttg cgggagatct
     1441 cagactacgc tctggaggga ctgcaaagcc tggagagtct ctctttctac gacaatcagc
     1501 tggcccaggt gcccaagcgg gcattggagc aggtgcctgg gctcaagttc ctagacctga
     1561 acaaaaaccc actgcagcgg gtcggaccgg gagactttgc caacatgctg cacctcaagg
     1621 aactaggact aaacaacatg gaggaactgg tctccatcga caagtttgct ctggtaaatc
     1681 tccctgagct gaccaagttg gacatcacca acaacccacg gctttctttc atccatcctc
     1741 gcgcctttca ccacctgccc cagatggaga ctctcatgct caacaacaat gctctcagtg
     1801 ccttgcacca gcagacagtg gagtctctgc ccaacctaca ggaggtgggg ctccacggca
     1861 accccatccg ctgtgactgt gtcatccgct gggccaacgc cacgggcacc cacgtccgtt
     1921 tcatcgaacc acagtccacc ctgtgtgctg agcctccaga ccttcagcgc cgcccagtcc
     1981 gggaggtgcc cttccgggag atgacagacc actgcctgcc cctcatctct ccccgaagtt
     2041 tcccctccag tctgcagata gccagtggag agagcacggt acttcactgt cgggcactgg
     2101 ctgaaccaga acctgagatc tactgggtca ccccagctgg agttcgactg acacctgctc
     2161 gctcaggcag gaggtaccga gtgttccctg agggtaccct agagttgcgg agggtgacgg
     2221 cggaagaggc aggcctatac acctgtgtgg cccagaactt ggtaggggct gacactaaaa
     2281 ccgtcagtgt tgtggttggc cacgctccct tccagccagg cagggacaaa ggaccaggac
     2341 tgaagctcca tgtgcaggag acacacccat atcacatcct gctgttttgg gcacccccac
     2401 ccaatatagt ctccaccaac ctcacctggt ctagcgcctc ctccctccgg gaccatgaag
     2461 ctcctgcttt ggcccgactg ccgcggggca cccaccgcta caacattacc cgcctccttc
     2521 cggccacaga gtactgggcc tgcctgcaag tggcctttgc tgatgcccac acccagttgg
     2581 cttgtgtttg ggccaggact aaagaggcct ctccttgcca cagagccctg ggggaccggc
     2641 ccgggctcat agccatcctg gctcttgcgg tcctcctgct ggcagctggg cttgcagccc
     2701 acctcggcag aggccagtcc aagcaggggg tggtgggtga gaagcctctt cttccggtct
     2761 gggctttctg gggctggagc gccccctctg tccgggtagt gtctgcaccc cttgtcctgc
     2821 cctggaatcc agggaggaag cagcccagat gtcagatggg gagagggtgc caccaccatt
     2881 gtctcaacat tgctgaagtt ccgccaggtc tcagcagtag agaaataact aggacaactc
     2941 tacacggatc cgactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca
     3001 aggatgacga tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa
     3061 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3121 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3181 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3241 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3301 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3361 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3421 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     3481 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3541 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3601 tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg
     3661 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     3721 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     3781 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     3841 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     3901 tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3961 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4021 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4081 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     4141 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4201 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4261 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     4321 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4381 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4441 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4501 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4561 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4621 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4681 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4741 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4801 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4861 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4921 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4981 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5041 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5101 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5161 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5221 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5281 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5341 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5401 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5461 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5521 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5581 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5641 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5701 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5761 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5821 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5881 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5941 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6001 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6061 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6121 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6181 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6241 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6301 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6361 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6421 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6481 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     6541 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6601 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6661 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6721 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6781 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6841 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6901 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6961 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7021 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7081 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7141 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7201 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7261 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7321 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7381 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7441 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7501 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7561 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7621 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7681 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7741 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7801 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7861 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7921 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7981 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8041 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     8101 gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     8161 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     8221 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
