LOCUS       V022072                 8467 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022072
VERSION     V022072
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..3987
                     /label="MERTK(NM_006343)"
                     /note="MERTK(NM_006343)"
                     /gene="MERTK"
     polyA_signal    4067..4291
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4337..4765
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4779..5108
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5175..5969
                     /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    6143..6264
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6313..6329)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6337..6353
                     /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(6361..6391)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6406..6427
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6715..7300)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7471..8331)
                     /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(8332..8436)
                     /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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atggggccgg ccccgctgcc gctgctgctg
     1021 ggcctcttcc tccccgcgct ctggcgtaga gctatcactg aggcaaggga agaagccaag
     1081 ccttacccgc tattcccggg accttttcca gggagcctgc aaactgacca cacaccgctg
     1141 ttatcccttc ctcacgccag tgggtaccag cctgccttga tgttttcacc aacccagcct
     1201 ggaagaccac atacaggaaa cgtagccatt ccccaggtga cctctgtcga atcaaagccc
     1261 ctaccgcctc ttgccttcaa acacacagtt ggacacataa tactttctga acataaaggt
     1321 gtcaaattta attgctcaat cagtgtacct aatatatacc aggacaccac aatttcttgg
     1381 tggaaagatg ggaaggaatt gcttggggca catcatgcaa ttacacagtt ttatccagat
     1441 gatgaagtta cagcaataat cgcttccttc agcataacca gtgtgcagcg ttcagacaat
     1501 gggtcgtata tctgtaagat gaaaataaac aatgaagaga tcgtgtctga tcccatctac
     1561 atcgaagtac aaggacttcc tcactttact aagcagcctg agagcatgaa tgtcaccaga
     1621 aacacagcct tcaacctcac ctgtcaggct gtgggcccgc ctgagcccgt caacattttc
     1681 tgggttcaaa acagtagccg tgttaacgaa cagcctgaaa aatccccctc cgtgctaact
     1741 gttccaggcc tgacggagat ggcggtcttc agttgtgagg cccacaatga caaagggctg
     1801 accgtgtcca agggagtgca gatcaacatc aaagcaattc cctccccacc aactgaagtc
     1861 agcatccgta acagcactgc acacagcatt ctgatctcct gggttcctgg ttttgatgga
     1921 tactccccgt tcaggaattg cagcattcag gtcaaggaag ctgatccgct gagtaatggc
     1981 tcagtcatga tttttaacac ctctgcctta ccacatctgt accaaatcaa gcagctgcaa
     2041 gccctggcta attacagcat tggtgtttcc tgcatgaatg aaataggctg gtctgcagtg
     2101 agcccttgga ttctagccag cacgactgaa ggagccccat cagtagcacc tttaaatgtc
     2161 actgtgtttc tgaatgaatc tagtgataat gtggacatca gatggatgaa gcctccgact
     2221 aagcagcagg atggagaact ggtgggctac cggatatccc acgtgtggca gagtgcaggg
     2281 atttccaaag agctcttgga ggaagttggc cagaatggca gccgagctcg gatctctgtt
     2341 caagtccaca atgctacgtg cacagtgagg attgcagccg tcaccagagg gggagttggg
     2401 cccttcagtg atccagtgaa aatatttatc cctgcacacg gttgggtaga ttatgccccc
     2461 tcttcaactc cggcgcctgg caacgcagat cctgtgctca tcatctttgg ctgcttttgt
     2521 ggatttattt tgattgggtt gattttatac atctccttgg ccatcagaaa aagagtccag
     2581 gagacaaagt ttgggaatgc attcacagag gaggattctg aattagtggt gaattatata
     2641 gcaaagaaat ccttctgtcg gcgagccatt gaacttacct tacatagctt gggagtcagt
     2701 gaggaactac aaaataaact agaagatgtt gtgattgaca ggaatcttct aattcttgga
     2761 aaaattctgg gtgaaggaga gtttgggtct gtaatggaag gaaatcttaa gcaggaagat
     2821 gggacctctc tgaaagtggc agtgaagacc atgaagttgg acaactcttc acagcgggag
     2881 atcgaggagt ttctcagtga ggcagcgtgc atgaaagact tcagccaccc aaatgtcatt
     2941 cgacttctag gtgtgtgtat agaaatgagc tctcaaggca tcccaaagcc catggtaatt
     3001 ttacccttca tgaaatacgg ggacctgcat acttacttac tttattcccg attggagaca
     3061 ggaccaaagc atattcctct gcagacacta ttgaagttca tggtggatat tgccctggga
     3121 atggagtatc tgagcaacag gaattttctt catcgagatt tagctgctcg aaactgcatg
     3181 ttgcgagatg acatgactgt ctgtgttgcg gacttcggcc tctctaagaa gatttacagt
     3241 ggcgattatt accgccaagg ccgcattgct aagatgcctg ttaaatggat cgccatagaa
     3301 agtcttgcag accgagtcta cacaagtaaa agtgatgtgt gggcatttgg cgtgaccatg
     3361 tgggaaatag ctacgcgggg aatgactccc tatcctgggg tccagaacca tgagatgtat
     3421 gactatcttc tccatggcca caggttgaag cagcccgaag actgcctgga tgaactgtat
     3481 gaaataatgt actcttgctg gagaaccgat cccttagacc gccccacctt ttcagtattg
     3541 aggctgcagc tagaaaaact cttagaaagt ttgcctgacg ttcggaacca agcagacgtt
     3601 atttacgtca atacacagtt gctggagagc tctgagggcc tggcccaggg ctccaccctt
     3661 gctccactgg acttgaacat cgaccctgac tctataattg cctcctgcac tccccgcgct
     3721 gccatcagtg tggtcacagc agaagttcat gacagcaaac ctcatgaagg acggtacatc
     3781 ctgaatgggg gcagtgagga atgggaagat ctgacttctg ccccctctgc tgcagtcaca
     3841 gctgaaaaga acagtgtttt accgggggag agacttgtta ggaatggggt ctcctggtcc
     3901 cattcgagca tgctgccctt gggaagctca ttgcccgatg aacttttgtt tgctgacgac
     3961 tcctcagaag gctcagaagt cctgatgtga gaattctgca gatatccagc acagtggcgg
     4021 ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     4081 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     4141 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     4201 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     4261 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     4321 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4381 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4441 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4501 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4561 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4621 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4681 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4741 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4801 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4861 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4921 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4981 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5041 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5101 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     5161 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5221 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     5281 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     5341 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5401 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5461 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5521 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5581 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5641 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5701 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5761 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5821 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5881 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5941 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     6001 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     6061 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     6121 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     6181 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6241 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     6301 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     6361 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     6421 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6481 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6541 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6601 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6661 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6721 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6781 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6841 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6901 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6961 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7021 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7081 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7141 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7201 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     7261 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7321 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7381 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7441 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7501 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7561 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7621 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7681 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7741 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7801 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7861 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7921 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7981 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8041 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8101 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8161 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8221 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8281 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8341 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8401 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8461 tgacgtc
//
