LOCUS       V020834                 8996 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020834
VERSION     V020834
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..3676
                     /label="MAMLD1(NM_001177465)"
                     /note="MAMLD1(NM_001177465)"
                     /gene="MAMLD1"
     CDS             3683..3748
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3772..4360
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4426..4474
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4676..5104
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5118..5447
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5514..6308
                     /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    6484..6605
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6654..6670)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6678..6694
                     /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(6702..6732)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6747..6768
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7056..7644)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7815..8675)
                     /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(8676..8780)
                     /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 ccatggatga ctggaaaagt cggcttgtaa tcaagagcat
      721 gcttccccat ttcgccatgg tgggaaatcg tcaggagccc agaaagctcc aggaatcggg
      781 aactgttaag aggagacaag aagaagacca cttccagttt ccagacatgg ctgatggggg
      841 ctaccctaat aaaattaaga ggccttgcct tgaagatgtc acccttgcaa tgggcccagg
      901 tgctcatcct agtactgctt gtgcagaact gcaggtccct ccattgacaa taaatcctag
      961 ccctgcggct atgggagtgg ctggccagtc attactgctg gagaataacc ctatgaatgg
     1021 caacatcatg ggctcaccat ttgtagtacc acagactaca gaagtgggac tgaaagggcc
     1081 cactgttcct tactatgaga aaatcaacag cgtgccggct gtagaccagg agcttcaaga
     1141 gctgctagag gagctcacca aaattcaaga cccttctcca aatgagctag atcttgagaa
     1201 gatactgggg acgaagccag aagagccact ggttttagat catccccagg caaccctaag
     1261 cacaactccc aagccttcgg ttcagatgtc acacttggag agcctggctt ccagcaagga
     1321 gtttgcttct agttgcagcc aagttactgg catgtcactt cagatcccat cctcctccac
     1381 agggatcagc tattcgattc cttccaccag taagcagata gtgtcaccga gttcttcaat
     1441 ggcacagtcc aagagccagg tccaggccat gctccctgtc gctctgcccc ccttaccagt
     1501 gcctcagtgg catcacgccc accagctgaa ggcgttggca gccagcaagc aggggtctgc
     1561 tacaaagcag caagggccca cccccagttg gtctggtctg cctcctccag gactctctcc
     1621 accttaccgc ccagtgccat caccacaccc accaccgctg ccactgccac caccaccacc
     1681 cccattcagc ccccagagcc tcatggtgtc ctgcatgtcg tccaatacct tgtcgggtag
     1741 cactctccga ggctctccca atgccttact gtcaagcatg acgtccagca gcaatgctgc
     1801 cctggggccc gccatgccct atgctcctga gaagctcccc agccctgctc tcactcaaca
     1861 gccgcagttc ggccctcaga gctccattct tgccaacctc atgtcctcta ccatcaaaac
     1921 ccctcaagga cacctgatgt ctgctctgcc tgccagcaac cctgggccgt ccccacccta
     1981 tcgcccagag aagctctcta gcccaggctt gccacagcag tccttcaccc cacagtgttc
     2041 cctgatccga agcctcactc ccaccagtaa tcttctaagc cagcaacagc agcagcagca
     2101 gcagcagcag caagcaaatg tgatctttaa gcccataagc agcaactcat ccaaaaccct
     2161 gagcatgatc atgcagcagg ggatggcaag ctccagccca ggagccacgg agccatttac
     2221 ttttggcaac accaagccct tgtcccattt tgtttctgag ccgggtcccc agaagatgcc
     2281 ctccatgcct accacctcta ggcagccttc cctgctccac tacctgcagc agccgacacc
     2341 aacgcaggcc tcctcagcca ctgcctcctc cacggccact gccaccttgc agctgcagca
     2401 gcagcagcag caacagcagc agcagcctga ccattcttca ttccttctgc agcagatgat
     2461 gcagcaaccc cagcgttttc agcgatcagt ggcctcagat tccatgcctg ctctgcccag
     2521 acagcaagag gaacagagat caggtcttat ggcaatgacc cctgaacggc agaatgcata
     2581 tatctcccaa cagatgagtc catttgaagc cgtccaagaa caagtcacct ccaagtgtag
     2641 ccggatcaag gcaagccccc catctagcaa gcacttgatg ccacccagaa ctgggcttct
     2701 tcagaacaat ctgagtccag gaatgatccc actcaccagg caccagagct gcgagggcat
     2761 gggagtgatc tcaccaactc tggggaagcg gcaaggaatt ttcacctcca gcccccagtg
     2821 tcccatcctc tcacactcag gccagactcc cctgggcaga cttgactctg tctgccagca
     2881 tatgcagagt cccaaggcca ccccaccaga agtgcccctg cctgggttct gtcccagctc
     2941 cctgggcacc cagtccttga gtccccacca gctcagacgg cctagtgtgc caagaatgcc
     3001 cactgcgttc aacaatgctg catgggtcac agcggcagca gctgtgacca cagcagtttc
     3061 ggggaaaaca cccctcagcc aagtggataa tagcgttcag cagcactcac cttctggcca
     3121 ggcctgcctt cagaggccat ctgattggga ggcacaagtg cccgctgcga tgggaacaca
     3181 agtgcccctg gccaacaacc ccagcttcag cctgctgggc agccagagcc tcaggcagag
     3241 cccggtacag ggcccggtgc ctgtagcaaa caccaccaag ttcctccagc agggtatggc
     3301 cagctttagt cccctgagcc ccatacaggg catcgagcca ccaagctatg tggctgctgc
     3361 tgccaccgct gctgctgctt ctgccgttgc tgccagccag ttcccaggtc cgttcgacag
     3421 aacggatatt ccccctgagc tgccacctgc cgactttttg cgccagcccc aacccccact
     3481 aaatgatctg atttcgtcac ctgactgcaa tgaggtagat ttcattgaag ctctcttgaa
     3541 aggctcctgt gtgagcccag atgaagactg ggtgtgcaac ttgaggctga tcgacgacat
     3601 tttggaacag catgctgctg ctcaaaatgc cacagcccag aattctgggc aagtcaccca
     3661 ggatgctggg gcacttctcg aggactacaa agaccatgac ggtgattata aagatcatga
     3721 catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga caatcaacct
     3781 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     3841 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     3901 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     3961 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     4021 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     4081 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     4141 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     4201 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     4261 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     4321 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa
     4381 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     4441 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     4501 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     4561 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     4621 gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg
     4681 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4741 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4801 gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct
     4861 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4921 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4981 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     5041 attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5101 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     5161 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     5221 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     5281 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     5341 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     5401 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     5461 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     5521 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     5581 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     5641 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     5701 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5761 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5821 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5881 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5941 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     6001 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     6061 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     6121 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     6181 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     6241 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     6301 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca
     6361 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     6421 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     6481 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     6541 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     6601 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     6661 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     6721 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     6781 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     6841 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     6901 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     6961 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     7021 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     7081 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     7141 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7201 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc
     7261 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     7321 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7381 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7441 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     7501 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7561 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7621 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     7681 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7741 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     7801 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7861 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7921 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7981 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     8041 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     8101 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     8161 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     8221 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     8281 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     8341 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     8401 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     8461 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     8521 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     8581 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     8641 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     8701 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     8761 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     8821 ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt
     8881 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     8941 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg
//
