LOCUS       V047246                 7317 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047246
VERSION     V047246
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..1997
                     /label="Rilpl1(NM_001417895)"
                     /note="Rilpl1(NM_001417895)"
                     /gene="Rilpl1"
     CDS             2004..2069
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2093..2681
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2747..2795
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2997..3425
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3439..3768
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3835..4629
                     /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    4805..4926
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4975..4991)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4999..5015
                     /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(5023..5053)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5068..5089
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5377..5965)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6136..6996)
                     /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(6997..7101)
                     /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 ccaccatgga ggagccgcta gggtcaccgc
      781 ccgccgccct gtcggctctg gagaagaacg tggcggagct gaccgtcatg gatgtgtacg
      841 atatcgcgtc gctcgtgggc cacgagttcg agagggtcat cgaccaacac ggctgcgagt
      901 ccatcgcacg cctcatgccc aaggtcgtga gggtcctgga gatcctcgag gtgctggtta
      961 gccggcacca cgtcgctccc gagctagacg agctgcgcct ggaactggac cggttgcgcg
     1021 tagagaggat ggaccgcatt gaaaaggagc gcaagcatca aaaggagctg gagctggtgg
     1081 aggacgtgtg gcggggtgaa gcacaggacc tcctgtccca gattgcccag ctgcaggagg
     1141 agaataagca gctgatgacc aacctcaacc acaaggacgt gggcttctcc gaggaggagt
     1201 tccagaagca ggaaggcatg tcagagcgtg agcggcaggt gatgaagagg ctgaaggagg
     1261 tggtggacaa gcagagggat gagctccgag ccaaagacag ggagctgggc ctgaagaatg
     1321 aggacgtcga ggctttgcag cagcagcaga cgaggctaat gaagatcaac catgaccttc
     1381 ggcaccgggt caccgtggtt gaggcccagg ggaaagccct gattgagcag aaggtggagc
     1441 tcgaagcaga cctgcagacc aaggagcagg agatgggcag cctgcgggct gaactcggaa
     1501 agcttcgcga gaggctgcag ggcgagcaca gccagaatgg ggaggaggag gaggctgaga
     1561 tccagccgca gccagatgga gaggaatcga tttcagatgc agagaaggcc gccctggacc
     1621 tcaaggaccc caaccgcccg aggttcacgc tccaggagct gcgggacgtc ctgcacgaga
     1681 ggaacgagct caagtccaag gtgtttctgc tgcaagagga actggcctac tataagagtg
     1741 aagaaataga agaagagaat cgaatacccc agcctccgcc cattacacac ccgaggacgt
     1801 ccccccagcc tgaatcgggg atcaagcgac ttaaagaagg tcctttgagt gacgggttta
     1861 gctttttctc ccgggacaag aagcgcctgg ccaacacgca gagacccaca cacatccacg
     1921 agtcctttgg ccagtgggca attacccagc gagatgacgg gtacacagag caaggacagg
     1981 aagccctgca gcacctggga tccgactaca aagaccatga cggtgattat aaagatcatg
     2041 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     2101 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2161 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2221 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     2281 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2341 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     2401 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2461 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     2521 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     2581 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     2641 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     2701 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     2761 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     2821 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     2881 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     2941 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     3001 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3061 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3121 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     3181 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3241 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3301 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3361 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3421 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3481 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3541 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3601 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3661 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3721 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3781 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3841 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3901 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3961 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4021 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4081 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4141 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4201 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4261 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4321 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4381 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4441 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4501 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4561 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4621 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     4681 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4741 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4801 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4861 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4921 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4981 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5041 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5101 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5161 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5221 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5281 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5341 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5401 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5461 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5521 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     5581 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5641 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5701 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5761 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     5821 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5881 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5941 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6001 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6061 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6121 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6181 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6241 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6301 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6361 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6421 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6481 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6541 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6601 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6661 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6721 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6781 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6841 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6901 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6961 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7021 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7081 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     7141 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     7201 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     7261 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
