LOCUS       V034689                 7774 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034689
VERSION     V034689
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..3264
                     /label="NSUN2(NM_017755)"
                     /note="NSUN2(NM_017755)"
                     /gene="NSUN2"
     CDS             3271..3336
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3374..3598
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3644..4072
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4086..4415
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4482..5276
                     /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    5450..5571
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5620..5636)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5644..5660
                     /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(5668..5698)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5713..5734
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6022..6607)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6778..7638)
                     /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(7639..7743)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggggc ggcggtcgcg gggtcggcgg ctccagcaac agcagcggcc ggaggacgcg
     1021 gaggatggcg ccgagggtgg tggaaagcgc ggcgaggcgg gctgggaagg aggctacccc
     1081 gagatcgtca aggagaacaa gctgttcgag cactactacc aggagctcaa gatcgtgccc
     1141 gagggcgagt ggggccagtt catggacgct ctcagggagc cgctcccggc cactttaaga
     1201 attactggtt acaaaagcca cgcaaaagag attctccatt gcttaaagaa caaatatttt
     1261 aaggaattgg aggacctgga ggtggacggt cagaaagttg aagttccaca gccactgagt
     1321 tggtatcctg aagaacttgc ctggcacaca aatttaagtc gaaaaatctt gagaaaatcg
     1381 ccacacttgg aaaagtttca tcagtttcta gttagtgaaa cagaatctgg aaatattagt
     1441 cgtcaagaag ctgttagcat gatcccacca ctgctcctca acgtgcggcc tcatcataag
     1501 atcttagata tgtgtgcagc acctggctca aagaccacac agttaattga aatgctacat
     1561 gccgacatga atgtcccctt tccagaggga tttgttattg cgaatgatgt ggacaacaag
     1621 cgctgctacc tgctcgtcca tcaagccaag aggctgagca gcccctgcat catggtggtc
     1681 aaccatgatg cctccagcat acccaggctc cagatagatg tggacggcag gaaagagatc
     1741 ctcttctatg atcgaatttt atgtgatgtc ccttgcagtg gagacggcac tatgagaaaa
     1801 aacattgatg tttggaaaaa gtggaccacc ttaaatagct tgcagctaca tggcttacag
     1861 ctgcggattg caacacgcgg ggctgaacag ctggctgaag gtggaaggat ggtgtattcc
     1921 acgtgttcac taaaccctat tgaggatgaa gcagtcatag catctttact ggaaaaaagt
     1981 gaaggtgctt tggagcttgc tgatgtgtct aatgaactgc cagggctgaa gtggatgcct
     2041 ggaatcacac agtggaaggt aatgacgaaa gatgggcagt ggtttacaga ctgggacgct
     2101 gttcctcaca gcagacacac ccagatccga cctaccatgt tccctccgaa ggacccagaa
     2161 aagctgcagg ccatgcacct ggagcgatgc cttaggatat taccccatca tcagaatact
     2221 ggagggtttt ttgtggcagt attggtgaaa aaatcttcaa tgccgtggaa taaacgtcag
     2281 ccaaagcttc agggtaaatc tgcagagacc agagaaagca cacagctgag ccctgcagat
     2341 ctcacagaag ggaaacccac agatccctct aagctggaaa gtccgtcatt cacaggaact
     2401 ggtgacacag aaatagctca tgcaactgag gatttagaga ataatggcag taagaaagat
     2461 ggcgtgtgtg gtcctcctcc atcaaagaaa atgaagttat ttggatttaa agaagatcca
     2521 tttgtattta ttcctgaaga tgacccatta tttccaccta ttgagaaatt ttatgctttg
     2581 gatccttcat tcccaaggat gaatttgtta actcggacta cagaagggaa gaaaaggcag
     2641 ctctacatgg tttctaagga gttgcggaat gtgctgctga ataacagtga gaagatgaag
     2701 gttattaaca cggggatcaa agtctggtgt agaaataaca gcggtgaaga gtttgactgt
     2761 gctttccggc tggcacagga gggaatatat acattgtatc catttattaa ctcaagaatt
     2821 attactgtat caatggaaga tgttaagata ctgttgaccc aggaaaatcc cttttttaga
     2881 aaactcagca gtgagaccta cagtcaagca aaggacctgg caaagggaag catcgtgctg
     2941 aagtatgaac cagattctgc gaatccagac gctctgcagt gtcccatcgt cttatgcgga
     3001 tggcggggaa aggcctccat tcgaactttt gtgcccaaga atgaacggct tcattatctc
     3061 aggatgatgg ggctggaggt attgggagaa aagaagaagg aaggggttat cctcacaaat
     3121 gagagtgcag ccagcaccgg acagccagac aatgacgtga ctgagggaca gagagcagga
     3181 gagcccaaca gcccagatgc agaagaggcc aacagtccag acgtgacagc aggctgtgac
     3241 ccggcggggg tccatccacc ccggctcgag gactacaaag accatgacgg tgattataaa
     3301 gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt ttaaacccgc
     3361 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3421 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3481 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3541 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3601 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3661 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3721 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3781 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3841 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3901 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3961 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     4021 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     4081 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4141 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4201 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4261 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4321 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4381 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4441 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4501 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4561 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4621 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4681 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4741 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4801 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4861 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4921 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4981 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     5041 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5101 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5161 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5221 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5281 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5341 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5401 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5461 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5521 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5581 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5641 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5701 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5761 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5821 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5881 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5941 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6001 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6061 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6121 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6181 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6241 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6301 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6361 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6421 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6481 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6541 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6601 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6661 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6721 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6781 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6841 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6901 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6961 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7021 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7081 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7141 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7201 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7261 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7321 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7381 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7441 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7501 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7561 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7621 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7681 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7741 gcgcacattt ccccgaaaag tgccacctga cgtc
//
