LOCUS       V024947                 6717 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024947
VERSION     V024947
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..1397
                     /label="KDELR3(NM_006855)"
                     /note="KDELR3(NM_006855)"
                     /gene="KDELR3"
     CDS             1404..1469
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1493..2081
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2147..2195
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2397..2825
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2839..3168
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3235..4029
                     /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    4205..4326
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4375..4391)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4399..4415
                     /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(4423..4453)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4468..4489
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4777..5365)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5536..6396)
                     /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(6397..6501)
                     /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 ccaccatgaa cgtgttccga atcctcggcg
      781 acctgagcca cctcctggcc atgatcttgc tgctggggaa gatctggagg tccaagtgct
      841 gcaagggcat ctctgggaag agccagatcc tgtttgctct cgtcttcacc accaggtacc
      901 tggacctgtt caccaacttc atctccatct acaacacagt aatgaaggtg gtttttctcc
      961 tctgtgccta tgttacagtg tacatgatat atgggaaatt ccgtaaaact tttgacagtg
     1021 agaatgacac attccgcctg gagtttcttc tggtcccagt cattggcctt tccttccttg
     1081 aaaactacag tttcactctg ctggagatcc tctggacttt ctctatctat ctggaatcag
     1141 tggctatcct gccccagctc ttcatgatca gcaagactgg agaggctgag accataacta
     1201 ctcactacct gttctttctg ggtctgtacc gggcactcta cctggctaac tggatcaggc
     1261 ggtaccagac tgagaatttc tatgaccaaa ttgcagtcgt gtctggagta gtacaaacca
     1321 tcttctactg tgacttcttc tacttgtatg tgaccaaagt ccttaaggga aagaagttaa
     1381 gtcttccaat gccaatcgga tccgactaca aagaccatga cggtgattat aaagatcatg
     1441 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     1501 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     1561 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     1621 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     1681 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     1741 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     1801 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     1861 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     1921 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     1981 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     2041 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     2101 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     2161 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     2221 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     2281 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     2341 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     2401 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2461 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2521 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     2581 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2641 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2701 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2761 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2821 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2881 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2941 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3001 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3061 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3121 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3181 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3241 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3301 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3361 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3421 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3481 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3541 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3601 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3661 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3721 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     3781 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3841 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3901 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3961 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4021 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     4081 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4141 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4201 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4261 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4321 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4381 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4441 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4501 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4561 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4621 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4681 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4741 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4801 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4861 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4921 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     4981 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5041 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5101 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5161 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     5221 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5281 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5341 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5401 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5461 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5521 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5581 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5641 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5701 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5761 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5821 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5881 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5941 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6001 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6061 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6121 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6181 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6241 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6301 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6361 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6421 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6481 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     6541 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     6601 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     6661 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
