LOCUS       V024146                 6983 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024146
VERSION     V024146
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             779..1663
                     /label="Lilra5(NM_001081239)"
                     /note="Lilra5(NM_001081239)"
                     /gene="Lilra5"
     CDS             1670..1735
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1759..2347
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2413..2461
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2663..3091
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3105..3434
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3501..4295
                     /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    4471..4592
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4641..4657)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4665..4681
                     /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(4689..4719)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4734..4755
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5043..5631)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5802..6662)
                     /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(6663..6767)
                     /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 ctcggatcca ctagtccagt gtggtggaat tcgccaccat
      781 gaccttcgtc ttcacagctg tgctctgtct ggggctgaat ctgggccaag agacctccat
      841 gtcggaaggg aatccgcaca agccaaccct tagtgttcag ccaggattgg tggttgccaa
      901 agggaagcag gtgaccatct catgtgaggt gaccacaggg gcccgggaat accgtcttta
      961 taaagagggg ggtccacatc cttggcgcac acggaacact ccgaagacca caaacaaagc
     1021 tcagttcttg atcccatcaa ttgaacaacg atctgggggg atatatcgct gttactataa
     1081 gacccccact gggtggtcag agcacagtga ccctctagag cttgcagtga caggactcta
     1141 tagtaaaccc agcctgtcca cccagcccag caatgttgtg aattcagggg agactgtaac
     1201 ccttcagtgt gtctcaacac tgggatttaa caggtttgtt ctgaccaagg aaggagaaca
     1261 gaagtggtcc ttgatccagg aatcagagtt tataaactct acggggcaat tccagggcct
     1321 atttactgtg ggccccgtga cccccagtca aaggtggata ttcagatgtt acggctatca
     1381 tgtcaacagc ccccaggtgt ggtcggaacc cagcgatctc ttggaaatac atgtgtcaga
     1441 agccgatcag cccctcaggc catcaccaaa catatcagac ccaaagacag tctcacagcc
     1501 ccagaattac acaatggaga acctcatcag gatgggggct tcaattttgg tccttgtgct
     1561 tcttggcgtt cttctgttcg aagctcagca cagccaaagg cagacccaac atgcagctgg
     1621 gagagaaagc agtgcttctt tcatggtggc agacacctgg gaactcgagg actacaaaga
     1681 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg
     1741 agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     1801 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     1861 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     1921 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     1981 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     2041 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2101 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     2161 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     2221 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     2281 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     2341 tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     2401 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt
     2461 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     2521 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     2581 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc
     2641 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2701 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2761 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc
     2821 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2881 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2941 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3001 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat
     3061 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3121 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3181 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3241 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3301 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3361 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3421 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3481 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3541 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3601 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3661 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3721 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3781 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3841 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3901 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     3961 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4021 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4081 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4141 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4201 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4261 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg
     4321 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4381 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4441 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     4501 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4561 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     4621 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4681 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4741 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4801 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4861 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4921 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4981 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5041 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5101 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5161 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5221 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5281 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5341 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5401 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5461 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     5521 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5581 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5641 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5701 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5761 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5821 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5881 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5941 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6001 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6061 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6121 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6181 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6241 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6301 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6361 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6421 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6481 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6541 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6601 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6661 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6721 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6781 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tcgactctca
     6841 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     6901 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     6961 attgcatgaa gaatctgctt agg
//
