LOCUS       V033136                 9471 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033136
VERSION     V033136
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..4151
                     /label="SEL1L3(NM_015187)"
                     /note="SEL1L3(NM_015187)"
                     /gene="SEL1L3"
     CDS             4158..4223
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4247..4835
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4901..4949
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5151..5579
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5593..5922
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5989..6783
                     /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    6959..7080
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7129..7145)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7153..7169
                     /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(7177..7207)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7222..7243
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7531..8119)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8290..9150)
                     /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(9151..9255)
                     /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 ccaccatgca gcggcgcggc gcggggctcg
      781 ggtggccgcg gcagcagcag cagcaacccc cgccgctcgc ggtcggcccc cgggccgcag
      841 ccatggtccc gagtggcggc gtcccccagg gcctcggcgg ccgctctgcc tgcgcgctgc
      901 tcctgctctg ctacctgaat gttgtaccat ctttgggtag gcagacttcc ctgacgacat
      961 cagtgatacc caaagctgag cagagcgtgg cttacaaaga ctttatttat tttactgtct
     1021 ttgaaggaaa cgttcgcaac gtttctgaag tctcggttga gtatttatgc tctcagcctt
     1081 gtgttgtcaa tttggaagca gttgtttcat ctgagttcag aagtagcatt cccgtgtaca
     1141 aaaaaaggtg gaagaatgag aaacatcttc acaccagcag gacacaaata gtacatgtga
     1201 aatttccaag cattatggtt tacagagatg attatttcat cagacattcc atctctgtat
     1261 ctgcagtgat agtacgcgcc tggattactc acaaatacag tggcagagac tggaatgtta
     1321 aatgggagga aaacttgctc catgctgtag caaagaatta taccctcctg cagaccatcc
     1381 cgccttttga acgccctttc aaagatcatc aagtgtgcct tgagtggaac atgggttata
     1441 tttggaacct tcgggcaaac aggattccac agtgtcctct ggaaaatgat gtggttgccc
     1501 tgcttggctt tccttatgcc tccagtggag aaaacacagg cattgtcaag aagttcccga
     1561 ggtttcggaa ccgagagctg gaggccactc gacgccagag gatggattac ccagtgttta
     1621 ctgtttcatt gtggctttat ttactccatt attgcaaggc caacctctgt gggattctgt
     1681 actttgttga ctctaatgag atgtacggca caccttctgt atttcttacg gaagagggct
     1741 atttgcatat tcagatgcat cttgtcaaag gggaagacct tgctgtaaaa actaaattca
     1801 tcataccttt gaaggagtgg tttcgactgg atatctcttt taacggaggc cagatagtag
     1861 taaccactag cattggacag gatttgaaaa gctaccacaa tcagaccatt agcttccggg
     1921 aggatttcca ttataatgac acagctgggt acttcattat tggagggagc aggtatgtgg
     1981 ctggcattga agggtttttt ggacccctga agtactatcg ccttcgcagt ctgcaccccg
     2041 cccagatttt taatcccctc cttgagaagc aacttgctga acaaatcaag ttatattatg
     2101 aaaggtgtgc tgaggttcaa gaaatagtat ctgtgtatgc atctgcagca aagcacgggg
     2161 gcgagagaca agaagcatgc cacctccaca actcctacct ggacctccag cgcaggtatg
     2221 ggagaccctc gatgtgcaga gccttcccct gggagaagga gctgaaagac aaacacccca
     2281 gcttgttcca ggcattgctg gagatggatc tgctgaccgt gccaaggaac caaaatgaat
     2341 ctgtatcaga aatcggtggg aagatatttg agaaggctgt aaagagactc tctagcattg
     2401 atggtcttca ccaaattagc tctatcgtcc cctttctgac ggattccagc tgctgtggat
     2461 accataaagc atcctactac cttgcagtct tttatgagac tggattaaat gttcctcggg
     2521 atcagctgca gggcatgttg tatagtttgg ttggaggcca ggggagtgag aggctgtctt
     2581 caatgaatct tgggtataaa cactaccagg gtattgacaa ctaccccctg gactgggaac
     2641 tgtcgtatgc ctactacagc aacattgcca ccaagacacc ccttgaccag cacacactgc
     2701 aaggagatca ggcatatgtt gaaacaatta gactaaaaga tgatgaaata ctcaaggtac
     2761 aaaccaaaga agatggagat gtctttatgt ggttgaagca tgaagctacc cgaggcaatg
     2821 cagcagctca gcaacgattg gcccagatgc tgttctgggg gcagcaaggt gtggccaaga
     2881 atcccgaagc agcaattgag tggtacgcca agggcgccct ggagacggag gatcctgcgt
     2941 taatctatga ctatgccatt gtgctattca agggtcaagg agtaaaaaag aacagacggc
     3001 ttgccttaga gctgatgaag aaagcagctt ccaagggatt gcatcaggca gtcaatggcc
     3061 tgggatggta ttaccacaaa ttcaagaaaa attacgccaa agcagcaaag tactggttaa
     3121 aagcagaaga aatggggaac ccagatgcgt catacaatct tggagtcctg catttggatg
     3181 gcatcttccc tggagttcct ggaaggaatc aaactttagc tggtgaatat ttccataagg
     3241 ctgcgcaagg tggacacatg gaagggacct tgtggtgttc tctctactat atcacaggca
     3301 acctggagac attccctaga gatcctgaga aagctgttgt atgggcaaaa catgtagctg
     3361 agaaaaatgg ctacttgggc catgtcatcc gcaaaggcct caatgcctac ctggaaggtt
     3421 catggcatga agctttgctg tattatgttt tagcagcaga aactggaatt gaagtgtcac
     3481 agacaaattt agcacacatc tgtgaggaga ggccagacct ggccaggaga tacttgggtg
     3541 ttaactgtgt ttggagatac tataatttct ctgtttttca aatcgatgct ccttcctttg
     3601 catatttgaa gatgggagac ctttactact atggccacca aaaccagtca caagacctgg
     3661 agttgtctgt gcagatgtac gcccaagccg ccctggatgg agactcccag ggatttttta
     3721 acctggccct gctaatcgag gaaggtacga taatcccaca ccatatcttg gatttcttgg
     3781 aaattgactc aactctccat tctaataaca tctccattct ccaggaactg tacgaaaggt
     3841 gctggagcca cagtaacgag gagtccttca gcccctgctc cttggcctgg ctttacctgc
     3901 acttgcggct tctctggggt gctatcctgc actcagccct gatctacttt ctgggaacct
     3961 ttctgctatc catattgatc gcctggactg tgcagtattt ccagtctgtc tcagcaagcg
     4021 atccccctcc aagaccatcc caggcctccc cagacactgc cacgtccact gcaagtccag
     4081 ctgtgactcc agctgcagat gcctctgacc aagaccagcc cacagtaact aataacccgg
     4141 agccacgtgg gggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     4201 actacaagga tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga
     4261 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     4321 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     4381 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     4441 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     4501 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     4561 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     4621 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     4681 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     4741 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     4801 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     4861 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     4921 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     4981 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     5041 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     5101 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     5161 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5221 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5281 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     5341 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5401 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5461 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5521 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     5581 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     5641 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     5701 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     5761 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5821 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     5881 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     5941 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     6001 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     6061 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     6121 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     6181 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     6241 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     6301 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     6361 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     6421 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     6481 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     6541 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     6601 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     6661 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     6721 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     6781 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     6841 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     6901 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     6961 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     7021 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     7081 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     7141 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     7201 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     7261 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     7321 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     7381 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     7441 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     7501 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     7561 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     7621 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7681 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     7741 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7801 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7861 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7921 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     7981 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     8041 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     8101 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     8161 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     8221 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     8281 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     8341 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     8401 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     8461 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     8521 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     8581 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     8641 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8701 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8761 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8821 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8881 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8941 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     9001 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     9061 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     9121 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     9181 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     9241 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     9301 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     9361 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     9421 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
