LOCUS       V022572                 9407 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022572
VERSION     V022572
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"
     CDS             683..4030
                     /label="EIF2AK3(NM_004836)"
                     /note="EIF2AK3(NM_004836)"
                     /gene="EIF2AK3"
     CDS             4070..4159
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    4183..4771
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4837..4885
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5087..5515
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5529..5858
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5925..6719
                     /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    6895..7016
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7065..7081)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7089..7105
                     /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(7113..7143)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7158..7179
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7467..8055)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8226..9086)
                     /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(9087..9191)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggagcg cgccatcagc ccggggctgc tggtacgggc
      721 gctgctgctg ctgctgctgc tgctggggct cgcggcaagg acggtggccg cggggcgcgc
      781 ccgtggcctc ccagcgccga cggcggaggc ggcgttcggc ctcggggcgg ccgctgctcc
      841 cacctcagcg acgcgagtac cggcggcggg cgccgtggct gcggccgagg tgactgtgga
      901 ggacgctgag gcgctgccgg cagccgcggg agagcaggag cctcggggtc cggaaccaga
      961 cgatgagaca gagttgcgac cgcgcggcag gtcattagta attatcagca ctttagatgg
     1021 gagaattgct gccttggatc ctgaaaatca tggtaaaaag cagtgggatt tggatgtggg
     1081 atccggttcc ttggtgtcat ccagccttag caaaccagag gtatttggga ataagatgat
     1141 cattccttcc ctggatggag ccctcttcca gtgggaccaa gaccgtgaaa gcatggaaac
     1201 agttcctttc acagttgaat cacttcttga atcttcttat aaatttggag atgatgttgt
     1261 tttggttgga ggaaaatctc tgactacata tggactcagt gcatatagtg gaaaggtgag
     1321 gtatatctgt tcagctctgg gttgtcgcca atgggatagt gacgaaatgg aacaagagga
     1381 agacatcctg cttctacagc gtacccaaaa aactgttaga gctgtcggac ctcgcagtgg
     1441 caatgagaag tggaatttca gtgttggcca ctttgaactt cggtatattc cagacatgga
     1501 aacgagagcc ggatttattg aaagcacctt taagcccaat gagaacacag aagagtctaa
     1561 aattatttca gatgtggaag aacaggaagc tgccataatg gacatagtga taaaggtttc
     1621 ggttgctgac tggaaagtta tggcattcag taagaaggga ggacatctgg aatgggagta
     1681 ccagttttgt actccaattg catctgcctg gttacttaag gatgggaaag tcattcccat
     1741 cagtcttttt gatgatacaa gttatacatc taatgatgat gttttagaag atgaagaaga
     1801 cattgtagaa gctgccagag gagccacaga aaacagtgtt tacttgggaa tgtatagagg
     1861 ccagctgtat ctgcagtcat cagtcagaat ttcagaaaag tttccttcaa gtcccaaggc
     1921 tttggaatct gtcactaatg aaaacgcaat tattccttta ccaacaatca aatggaaacc
     1981 cttaattcat tctccttcca gaactcctgt cttggtagga tctgatgaat ttgacaaatg
     2041 tctcagtaat gataagtttt ctcatgaaga atatagtaat ggtgcacttt caatcttgca
     2101 gtatccatat gataatggtt attatctacc atactacaag agggagagga acaaacgaag
     2161 cacacagatt acagtcagat tcctcgacaa cccacattac aacaagaata tccgcaaaaa
     2221 ggatcctgtt cttcttttac actggtggaa agaaatagtt gcaacgattt tgttttgtat
     2281 catagcaaca acgtttattg tgcgcaggct tttccatcct catcctcaca ggcaaaggaa
     2341 ggagtctgaa actcagtgtc aaactgaaaa taaatatgat tctgtaagtg gtgaagccaa
     2401 tgacagtagc tggaatgaca taaaaaactc tggatatata tcacgatatc taactgattt
     2461 tgagccaatt caatgcctgg gacgtggtgg ctttggagtt gtttttgaag ctaaaaacaa
     2521 agtagatgac tgcaattatg ctatcaagag gatccgtctc cccaataggg aattggctcg
     2581 ggaaaaggta atgcgagaag ttaaagcctt agccaagctt gaacacccgg gcattgttag
     2641 atatttcaat gcctggctcg aagcaccacc agagaagtgg caagaaaaga tggatgaaat
     2701 ttggctgaaa gatgaaagca cagactggcc actcagctct cctagcccaa tggatgcacc
     2761 atcagttaaa atacgcagaa tggatccttt cgctacaaaa gaacatattg aaatcatagc
     2821 tccttcacca caaagaagca ggtctttttc agtagggatt tcctgtgacc agacaagttc
     2881 atctgagagc cagttctcac cactggaatt ctcaggaatg gaccatgagg acatcagtga
     2941 gtcagtggat gcagcataca acctccagga cagttgcctt acagactgtg atgtggaaga
     3001 tgggactatg gatggcaatg atgaggggca ctcctttgaa ctttgtcctt ctgaagcttc
     3061 tccttatgta aggtcaaggg agagaacctc ctcttcaata gtatttgaag attctggctg
     3121 tgataatgct tccagtaaag aagagccgaa aactaatcga ttgcatattg gcaaccattg
     3181 tgctaataaa ctaactgctt tcaagcccac cagtagcaaa tcttcttctg aagctacatt
     3241 gtctatttct cctccaagac caaccacttt aagtttagat ctcactaaaa acaccacaga
     3301 aaaactccag cccagttcac caaaggtgta tctttacatt caaatgcagc tgtgcagaaa
     3361 agaaaacctc aaagactgga tgaatggacg atgtaccata gaggagagag agaggagcgt
     3421 gtgtctgcac atcttcctgc agatcgcaga ggcagtggag tttcttcaca gtaaaggact
     3481 gatgcacagg gacctcaagc catccaacat attctttaca atggatgatg tggtcaaggt
     3541 tggagacttt gggttagtga ctgcaatgga ccaggatgag gaagagcaga cggttctgac
     3601 cccaatgcca gcttatgcca gacacacagg acaagtaggg accaaactgt atatgagccc
     3661 agagcagatt catggaaaca gctattctca taaagtggac atcttttctt taggcctgat
     3721 tctatttgaa ttgctgtatc cattcagcac tcagatggag agagtcagga ccttaactga
     3781 tgtaagaaat ctcaaatttc caccattatt tactcagaaa tatccttgtg agtacgtgat
     3841 ggttcaagac atgctctctc catcccccat ggaacgacct gaagctataa acatcattga
     3901 aaatgctgta tttgaggact tggactttcc aggaaaaaca gtgctcagac agaggtctcg
     3961 ctccttgagt tcatcgggaa caaaacattc aagacagtcc aacaactccc atagcccttt
     4021 gccaagcaat gaattctgca gatatccagc acagtggcgg ccgctcgagg aacaaaaact
     4081 aatatcagag gaagatttgg aacagaaact aatcagtgaa gaagatttag agcaaaagtt
     4141 aatttccgag gaggacttat agtgagggcc cgatatctcg acaatcaacc tctggattac
     4201 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     4261 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     4321 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     4381 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     4441 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     4501 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     4561 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
     4621 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     4681 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     4741 agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac
     4801 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     4861 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     4921 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     4981 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     5041 cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc
     5101 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     5161 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     5221 ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac
     5281 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     5341 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5401 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg
     5461 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     5521 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5581 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     5641 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5701 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5761 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     5821 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     5881 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     5941 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     6001 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     6061 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     6121 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     6181 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     6241 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     6301 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     6361 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     6421 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     6481 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     6541 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     6601 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     6661 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     6721 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     6781 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     6841 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     6901 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6961 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     7021 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     7081 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     7141 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     7201 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     7261 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7321 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7381 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7441 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7501 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7561 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7621 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     7681 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7741 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7801 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7861 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     7921 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7981 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     8041 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     8101 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     8161 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     8221 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     8281 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8341 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8401 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8461 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8521 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8581 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8641 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8701 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8761 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8821 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8881 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8941 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     9001 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     9061 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     9121 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     9181 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     9241 ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     9301 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     9361 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
