LOCUS       V023989                 7367 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023989
VERSION     V023989
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..2857
                     /label="ILDR2(NM_199351)"
                     /note="ILDR2(NM_199351)"
                     /gene="ILDR2"
     CDS             2864..2929
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2967..3191
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3237..3665
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3679..4008
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4075..4869
                     /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    5043..5164
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5213..5229)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5237..5253
                     /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(5261..5291)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5306..5327
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5615..6200)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6371..7231)
                     /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(7232..7336)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggataggg tcttgctgag
      961 gtggatttct ctcttctggc taacagccat ggtcgaaggc cttcaggtca cagtgcccga
     1021 caagaagaag gtggccatgc tcttccagcc cactgtgctt cgctgccact tctcaacatc
     1081 ctcccatcag cctgcagttg tgcagtggaa gttcaagtcc tactgccagg atcgcatggg
     1141 agaatccttg ggcatgtcct ctacccgggc ccaatctctc agcaagagaa acctggaatg
     1201 ggacccctac ttggattgtt tggacagcag gaggactgtt cgagtagtag cttcaaaaca
     1261 gggctcgact gtcaccctgg gagatttcta caggggcaga gagatcacga ttgttcatga
     1321 tgcagatctt caaattggaa agcttatgtg gggagacagc ggactctatt actgtattat
     1381 caccacccca gatgacctgg aggggaaaaa tgaggactca gtggaactgc tggtgttggg
     1441 caggacaggg ctgcttgctg atctcttgcc cagttttgct gtggagatta tgccagagtg
     1501 ggtgtttgtt ggcctggtgc tcctgggcgt cttcctcttc ttcgtcctgg tggggatctg
     1561 ctggtgccag tgctgccctc acagctgctg ctgctatgtc cgctgcccat gctgcccaga
     1621 ttcctgctgc tgccctcaag ccttgtatga agcagggaaa gcagcaaagg ccgggtaccc
     1681 tccctctgtc tccggtgtcc ccggccctta ctccatcccc tctgtccctt tgggaggagc
     1741 cccctcatct ggcatgctga tggacaagcc gcatccacct cccttggcac caagtgactc
     1801 cactggagga agccacagtg ttcgcaaagg ttaccggatc caggctgaca aagagagaga
     1861 ctccatgaag gtcctgtact atgttgagaa ggagctggct cagtttgatc cagccagaag
     1921 gatgagaggc agatataaca acaccatctc agaactcagc tccctacatg aggaggacag
     1981 caatttccgc cagtctttcc atcagatgag aagcaagcag ttccctgtgt ctggggactt
     2041 ggagagcaat cctgactatt ggtcaggtgt catgggaggc agcagtgggg caagccgcgg
     2101 gccctcagcc atggagtata acaaagagga tcgagagagc ttcaggcaca gccagccgcg
     2161 ctccaagtcg gagatgctgt cgcggaagaa cttcgccacg ggggtgccgg ccgtttccat
     2221 ggacgagctg gcggccttcg ctgactccta cggccagcgg ccccgccggg cagacggcaa
     2281 cagtcacgag gcgcggggcg ggagccgctt cgagcgctcg gagtcgcggg cgcacagcgg
     2341 cttctaccag gacgactcct tggaggagta ctacggtcag cgcagccgca gccgcgagcc
     2401 cctgaccgat gctgaccgcg gctgggcctt cagccccgcg cgccgcagac ccgccgagga
     2461 cgcgcacctg ccgcggctgg tgagccgcac gccaggcacc gcacccaaat acgaccactc
     2521 gtacctgggc agcgcgcggg agcgccaggc gcggcccgag ggcgccagcc gcggtggcag
     2581 cctggagacg ccatccaagc ggagcgcgca gctcggcccg cgcagcgcct cctactacgc
     2641 ttggtcgccg cccggcacct acaaggccgg ctcgtcgcag gacgaccagg aggacgcgtc
     2701 cgacgacgcg ctgccgccct acagcgagct ggagctgacc cgcggcccgt cctaccgcgg
     2761 ccgcgacctg ccctaccaca gcaactcgga gaagaagagg aaaaaggagc ccgccaagaa
     2821 aaccaatgac tttccaacca ggatgtccct tgtggtcctc gaggactaca aagaccatga
     2881 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc
     2941 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3001 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3061 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3121 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3181 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3241 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3301 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3361 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3421 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3481 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3541 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3601 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3661 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3721 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3781 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3841 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3901 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3961 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4021 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4081 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4141 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4201 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4261 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4321 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4381 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4441 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4501 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4561 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4621 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4681 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4741 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4801 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4861 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4921 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4981 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5041 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5101 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5161 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5221 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5281 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5341 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5401 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5461 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5521 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5581 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5641 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5701 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5761 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5821 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5881 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5941 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6001 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6061 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6121 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     6181 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6241 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6301 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6361 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6421 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6481 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6541 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6601 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6661 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6721 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6781 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6841 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6901 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6961 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7021 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7081 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7141 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7201 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7261 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7321 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
