LOCUS       V028868                 9936 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028868
VERSION     V028868
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..4616
                     /label="CDON(NM_001243597)"
                     /note="CDON(NM_001243597)"
                     /gene="CDON"
     CDS             4623..4688
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4712..5300
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5366..5414
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5616..6044
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6058..6387
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6454..7248
                     /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    7424..7545
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7594..7610)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7618..7634
                     /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(7642..7672)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7687..7708
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7996..8584)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8755..9615)
                     /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(9616..9720)
                     /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 tccggatctt ggacccttat
      781 gtacactgct gtatgttact cttacaattc tgtgctcttc tgtgagttca gacttggcac
      841 cttattttac ttctgagccg ctctctgctg tccagaaact tggtggacct gtagtactgc
      901 attgttctgc tcaacctgtg accactcgta tctcatggct gcataacgga aaaacattgg
      961 atggaaacct ggaacatgtt aagattcatc aggggactct gacaattctt tctctcaact
     1021 cctctctttt gggttactac cagtgccttg ccaacaatag catcggtgcc attgtgagtg
     1081 gccctgcgac agtatctgtg gcagttcttg gtgattttgg ttcatccaca aagcatgtta
     1141 ttacagcaga agaaaaaagt gctggtttca ttggctgcag ggtaccggag agtaacccca
     1201 aagctgaggt gcgctataaa atccggggaa aatggctgga acattccaca gagaattact
     1261 taatccttcc atcaggaaat cttcagattt tgaatgtatc cttagaggac aagggatcat
     1321 acaaatgtgc agcttataat cctgtcacac atcaattaaa agttgaacct attggccgaa
     1381 agctccttgt gagtcgtcct tcttcagatg atgttcacat tcttcacccc acccattcac
     1441 aggcattagc tgttctttct cgtagccctg taaccttgga gtgtgtggtg agtggggtcc
     1501 cggctcctca agtgtattgg ctaaaggacg ggcaggacat tgcaccagga agcaactgga
     1561 gaaggttgta ttctcatctt gccactgata gcgttgaccc ggcggactcc ggaaactatt
     1621 cctgcatggc gggaaacaag tctggagatg taaaatatgt gacttacatg gttaatgtac
     1681 ttgaacatgc ttccatttct aaaggactac aggatcagat agtgtctctg ggtgccacag
     1741 tacactttac ctgcgacgtt catgggaacc cagcccccaa ctgtacctgg tttcacaatg
     1801 cacagcctat tcatccttct gcacgacatc taactgcagg aaacggactg aaaatcagtg
     1861 gggttactgt ggaagatgtt gggatgtatc agtgtgtagc agataatggg attggattta
     1921 tgcactctac tggaagactt gaaattgaaa atgacggtgg attcaagcca gttataatta
     1981 cggcaccagt aagtgcaaag gttgcagacg gagactttgt tactctgtcc tgcaatgcca
     2041 gtgggctgcc ggttccggtc attcgttggt atgacagcca tggattgata accagccatc
     2101 catctcaagt cctgagatcg aaatcccgaa aatcacagtt atcaagacct gagggcttga
     2161 acctggagcc tgtgtacttc gtcctgtccc aagctggtgc aagctctctc catattcagg
     2221 ctgtgactca ggaacatgcg gggaaataca tctgcgaagc tgcaaatgaa catggtacca
     2281 cacaggcaga agcatctctc atggttgttc cttttgaaac aaatacaaaa gcagagacag
     2341 tcacacttcc tgatgctgct cagaatgatg acagaagtaa gagagatggt tcagaaactg
     2401 ggttactgag ctcatttccg gtgaaggtcc atcccagtgc agtggaatca gcaccagaga
     2461 aaaacgccag cggcatctct gttcctgatg cccccatcat actgagcccc ccacagaccc
     2521 acacaccaga cacgtacaac ctggtgtgga gggcaggcaa ggatggtggg ctgcccatca
     2581 atgcttactt tgtgaagtat cgaaagctgg atgatggggt tggcatgctg ggaagctggc
     2641 acacggttcg agtcccagga agtgaaaatg agctccattt agctgagctg gagccatcta
     2701 gtctttatga agtcttgatg gtagcaagaa gcgcagcagg tgaaggccaa cctgccatgc
     2761 ttaccttccg aaccagcaaa gaaaaaacag cgtcatcaaa aaacacccag gcatcctctc
     2821 cacccgtggg catccctaag tatcccgttg tttcagaggc tgcaaacaac aattttggag
     2881 tggtacttac agattcctct aggcacagtg gagttccaga ggcaccagat cggcctacca
     2941 tctccactgc atcagagaca tcagtctatg tcacttggat tcctcgggca aacgggggtt
     3001 ctccaatcac tgccttcaaa gtcgaatata aacggatgag gaccagcaat tggctggtgg
     3061 cagctgaaga catccctcct tccaaacttt cagtggaagt tcgtagttta gaaccaggtt
     3121 caacatacaa atttagggtc attgccatca accattatgg tgagagtttt cggagttcag
     3181 catctcgtcc ttatcaagtg gttgggttcc ccaatcgctt ttccagccgt ccaataactg
     3241 gacctcacat tgcatacaca gaggctgtca gcgatactca gatcatgcta aagtggacgt
     3301 acattccatc aagtaacaat aacactccca ttcaaggatt ttatatctat taccgaccaa
     3361 cagatagtga caatgacagt gattacaaga gggatgttgt agaaggttca aagcagtggc
     3421 acatgattgg ccacctgcag ccagaaacct cctatgacat taaaatgcaa tgcttcaatg
     3481 aaggaggaga aagtgaattt agcaatgtga tgatctgcga gactaaagtg aaacgtgttc
     3541 ctggagcttc tgaatatcct gtcaaagact tgagtacccc tccaaattct ttgggaagtg
     3601 gaggaaatgt ggggcctgca accagccctg ccagaagcag tgacatgtta tatctgatcg
     3661 ttggctgtgt gctgggcgtc atggtcctca ttctgatggt tttcattgca atgtgcctgt
     3721 ggaagaatcg ccagcagaat accatacaaa aatatgaccc accaggatat ctctaccaag
     3781 gatcagatat gaacgggcag atggtggact acaccactct ctcaggagca agtcagataa
     3841 atggaaatgt tcacggaggc ttcctaacca atggcggtct cagcagtggc tattcccacc
     3901 ttcaccataa ggtccccaat gcagtcaatg gaattgtgaa tgggagccta aatggagggc
     3961 tttactccgg gcacagcaac tctctaacca ggacacacgt ggattttgaa catcctcatc
     4021 atctagtgaa tggtggtgga atgtacacgg ccgtgcctca gattgaccct ctggagtgtg
     4081 ttaactgccg aaattgtcga aacaacaata ggtgtttcac caaaaccaac agcactttca
     4141 gcagcagccc tcctcctgtg gtccctgtgg tagcacctta tcctcaggat ggtttggaaa
     4201 tgaagcccct cagtcacgtg aaggtgcctg tatgcctgac ttccgcagtc cctgattgtg
     4261 gccagttgcc ggaggagagc gtcaaggaca atgtggaacc agtccctact cagcgtacct
     4321 gctgtcagga cattgtaaat gacgtcagct ctgatggctc agaagatcca gcagagttca
     4381 gcagaggtca ggaaggcatg atcaacctta ggattccaga tcatcttcag ttagctaaga
     4441 gctgtgtatg ggaaggagac agctgtgccc attcagaaac agagatcaac attgtaagtt
     4501 ggaatgctct tattttgcca cctgtccccg agggctgtgc tgagaagaca atgtggtctc
     4561 cacctggcat tcctttagac agcccgacag aggtccttca gcagccccgg gaaaccggat
     4621 ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg
     4681 atgacaagta gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga
     4741 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     4801 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     4861 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     4921 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     4981 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     5041 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     5101 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     5161 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     5221 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     5281 cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca
     5341 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
     5401 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     5461 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     5521 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     5581 agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     5641 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5701 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5761 tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5821 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5881 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5941 actcaaccct atctcggtct attcttttga tttataaggg attttgggga tttcggccta
     6001 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     6061 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     6121 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     6181 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     6241 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     6301 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     6361 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     6421 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     6481 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     6541 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     6601 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     6661 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     6721 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     6781 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     6841 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     6901 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     6961 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     7021 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     7081 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     7141 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     7201 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     7261 ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     7321 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     7381 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     7441 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     7501 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     7561 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     7621 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     7681 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     7741 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     7801 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     7861 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     7921 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     7981 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     8041 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     8101 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     8161 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     8221 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     8281 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     8341 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     8401 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     8461 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     8521 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     8581 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     8641 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     8701 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     8761 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     8821 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     8881 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     8941 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     9001 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     9061 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     9121 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     9181 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     9241 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     9301 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     9361 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     9421 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     9481 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     9541 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     9601 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     9661 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     9721 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     9781 tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     9841 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     9901 ggcttgaccg acaattgcat gaagaatctg cttagg
//
