LOCUS       V045987                 9183 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045987
VERSION     V045987
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..3863
                     /label="Kansl1(NM_001081045)"
                     /note="Kansl1(NM_001081045)"
                     /gene="Kansl1"
     CDS             3870..3935
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3959..4547
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4613..4661
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4863..5291
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5305..5634
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5701..6495
                     /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    6671..6792
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6841..6857)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6865..6881
                     /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(6889..6919)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6934..6955
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7243..7831)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8002..8862)
                     /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(8863..8967)
                     /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 ccaccatggc tgcgatggcg cccgctctca
      781 ctgacgcagc agctgaagca caccacatcc ggttcaaact ggctccccca tcctccacct
      841 tgtcccctgg cagtgccgaa aataacggca acgccaatat tcttatttct gccaacggaa
      901 ccaaaagaaa agccattgct gcggaggatc ccagtctaga cttccgcaat aatcctacta
      961 aggaagactt gggaaagctg cagccattgg tggcatctta tctctgctct gatgtaacat
     1021 ctgttcccgc aaaagagtct ttgaaattgc aaggcgtttt cagcaagcag acagtcctta
     1081 agtctcatcc tcttttatct cagtcctatg aactccgagc tgagctgctg gggaggcagc
     1141 cagttttgga gttttcctta gaaaacctta gaaccatgaa tacaagtggt cagacagctc
     1201 tgccacaagc gcctgtaaat ggattggcta agaaattgac taaaagttca acacattcgg
     1261 atcatgacaa ttccagttcc ctcaatgggg gaaaacggtc tctcacttcg tcatctctac
     1321 aaggtggtga ggtgggggga cctgactctg ggaacttgaa ggggggtatg accaattgca
     1381 ctcttccaca tagaagcctt gatatacaac acacaactct atatagcaat aatagcactg
     1441 caaacaaatc ttctgttaat tccatggacc agccggcact tcaaggaagc agcaggttat
     1501 cacctagtac agactccagc tctaacttga caaatgtgaa gttagaagtc aaaaagtctc
     1561 ctctgtcttc cattcttttc agtgctttag actctgatac aaggataaca gctttactga
     1621 ggcggcaggc tgatattgag atccgtgccc gaaggttaca gaagcgatta caggttgtgc
     1681 aagccaagca ggtggagaga catttacagc accaactggg tggatttttg gagacgacat
     1741 tgagtaaact gccaaacttg gaatccttaa ggtcccggag ccagttgatg ttgacccgaa
     1801 aggctgaagc tgccttgaga aaagctgcca gtgagtctgc cacttcagag ggacttagca
     1861 actttttgaa aagtgattca atttcagaag aattggagag atttacagct agtggcatag
     1921 ccaacctgag gtgcagtgag caagcgtttg attcggatgt cacagacagt agttccggag
     1981 gggagtctga cattgaagag gaagaactga ctcgagctga tcctgagcaa tgccatgtgc
     2041 ccctgaaacg taggtctgaa tggagatggg ctgcagaccg agcagctatt gtcagccgct
     2101 ggaactggct tcaggcccat gtttctgact tggaatatcg aattcgtcag caaacagata
     2161 tttacaaaca gatacgtgct aataaggggt tgatagtcct tggggaggca cctttcccag
     2221 atcatacaac agacttactt tcacttagtt ctgaggtgaa gacagatcat gggagagata
     2281 aattgattga gtctgtttct cagccatcag aaaaccatgg tatccttgtt agtaatatta
     2341 cagagtcatt gtctaccaaa tcatgtggag caccgcgacc tgtcaatgga gttgttaaca
     2401 gtcttcagcc tgtcctggca gaccaagttc caggtgacag ctctgatgct gaggaacaat
     2461 tacataaaaa gcaaaggctg aatttagttt cttcatctga tggcacgtgt gtcgcagccc
     2521 gaacccggcc tgtactgacc tgtaagaaac ggaggcttgt taggcccagc agcatcgttc
     2581 ctctttctaa gaaggttcac cgtaatgtcc gctctggctg tgatgtgaat ccctcttgtg
     2641 cactgtgtgg ttcaggcagt gtgaacacca tgcctcctga aatacactat gaagcacctc
     2701 tcttagagcg tctttcccag ttggactcct gcgttcaccc tgtgctagca tttccagatg
     2761 atgttcccac aagcctgcac ttccagagca tgctgaagtc tcagtggcaa aacaagcctt
     2821 ttgacaagat caaacctacc aaaaagtttt ctcttaagca cagagcgacc atgccatgca
     2881 gtctgtcaga tccagttcgt aaagacagac acaagttggt caactccttc cttacaacag
     2941 ccatgttgaa gcatcacaca gacatgagca gtccgagcta cttgacagct acccaccatc
     3001 caccccacag ccccctggtg agacagctct ccacctcatc agacacctcc acacccacca
     3061 gctcgggctc acaagttgca gccagcacat ctcagccagt gaggaggaga aggggagaga
     3121 gctcatttga tattaacaac attgtcatcc caatgtctgt tgctgcaaca actcgtgttg
     3181 agaaactaca atacaaagaa atccttactc caagctggcg agaggttgat gttcagtctc
     3241 tgaaggggag tcctgatgaa gagaatgagg agatcgagga cctatctgat gcagcctttg
     3301 cggccctgca tgccaaatgt gaggagatgg aaagggcaag gtggctgtgg accacaagtg
     3361 tgccacccca gcgtcggggc agcaggtctt acaggtcatc agacggccgg accacccctc
     3421 agttgggcag tgccaacccc tccaccccac agcctgcgtc ccctgatgtc agtagtagcc
     3481 actctctgtc agaattctcc catggtcaat ctcctaggag tcccattagc ccagaactac
     3541 actcagcccc tctcacccct gtggctcggg acagtctgcg acacctggcc agtgaagaca
     3601 cccgatgttc cacaccagag ctgggactag atgaacagtc agtccagcca tgggaacgac
     3661 ggaccttccc tctggcctac agtccccagg cagaatgtga ggagcagctg gatgcacagg
     3721 acacggcagc ccgctgcact cgccgcacct caggcagcaa gactggccgg gaggcagagg
     3781 tggcacccac ctcgcctcct gttgtcccgc tcaagagtcg gcatctggca gcaacagtca
     3841 cagctcagcg cccagctcac agaggatccg actacaaaga ccatgacggt gattataaag
     3901 atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa
     3961 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     4021 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     4081 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     4141 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     4201 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     4261 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     4321 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     4381 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     4441 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     4501 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg
     4561 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     4621 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     4681 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     4741 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     4801 cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc
     4861 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4921 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4981 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     5041 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5101 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5161 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5221 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5281 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     5341 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     5401 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5461 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5521 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     5581 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     5641 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     5701 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5761 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5821 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5881 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5941 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     6001 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     6061 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     6121 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     6181 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     6241 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6301 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6361 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     6421 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6481 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     6541 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     6601 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6661 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6721 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6781 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     6841 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6901 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6961 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     7021 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     7081 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     7141 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7201 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7261 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7321 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7381 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     7441 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7501 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7561 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7621 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7681 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7741 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7801 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7861 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7921 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7981 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     8041 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     8101 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     8161 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8221 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8281 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8341 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8401 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8461 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8521 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8581 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8641 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8701 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8761 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8821 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8881 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8941 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     9001 cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc
     9061 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     9121 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     9181 agg
//
