LOCUS       V024847                 8162 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024847
VERSION     V024847
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             941..3652
                     /label="DLG1(NM_001290983)"
                     /note="DLG1(NM_001290983)"
                     /gene="DLG1"
     CDS             3659..3724
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3762..3986
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4032..4460
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4474..4803
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4870..5664
                     /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    5838..5959
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6008..6024)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6032..6048
                     /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(6056..6086)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6101..6122
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6410..6995)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7166..8026)
                     /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(8027..8131)
                     /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 atccgccacc atgccggtcc ggaagcaaga
      961 tacccagaga gcattgcacc ttttggagga atatcgttca aaactaagcc aaactgaaga
     1021 cagacagctc agaagttcca tagaacgggt tattaacata tttcagagca acctctttca
     1081 ggctttaata gatattcaag aattttatga agtgacctta ctggataatc caaaatgtat
     1141 agatcgttca aagccgtctg aaccaattca acctgtgaat acttgggaga tttccagcct
     1201 tccaagctct actgtgactt cagagacact gccaagcagc cttagcccta gtgtagagaa
     1261 atacaggtat caggatgaag atacacctcc tcaagagcat atttccccac aaatcacaaa
     1321 tgaagtgata ggtccagaat tggttcatgt ctcagagaag aacttatcag agattgagaa
     1381 tgtccatgga tttgtttctc attctcatat ttcaccaata aagccaacag aagctgttct
     1441 tccctctcct cccactgtcc ctgtgatccc tgtcctgcca gtccctgctg agaatactgt
     1501 catcctaccc accataccac aggcaaatcc tcccccagta ctggtcaaca cagatagctt
     1561 ggaaacacca acttacgtta atggcacaga tgcagattat gaatatgaag aaatcacact
     1621 tgaaagggga aattcagggc ttggtttcag cattgcagga ggtacggaca acccacacat
     1681 tggagatgac tcaagtattt tcattaccaa aattatcaca gggggagcag ccgcccaaga
     1741 tggaagattg cgggtcaatg actgtatatt acgagtaaat gaagtagatg ttcgtgatgt
     1801 aacacatagc aaagcagttg aagcgttgaa agaagcaggg tctattgtac gcttgtatgt
     1861 aaaaagaagg aaaccagtgt cagaaaaaat aatggaaata aagctcatta aaggtcctaa
     1921 aggtcttggg tttagcattg ctggaggtgt tggaaatcag catattcctg gggataatag
     1981 catctatgta accaaaataa ttgaaggagg tgcagcacat aaggatggca aacttcagat
     2041 tggagataaa cttttagcag tgaataacgt atgtttagaa gaagttactc atgaagaagc
     2101 agtaactgcc ttaaagaaca catctgattt tgtttatttg aaagtggcaa aacccacaag
     2161 tatgtatatg aatgatggct atgcaccacc tgatatcacc aactcttctt ctcagcctgt
     2221 tgataaccat gttagcccat cttccttctt gggccagaca ccagcatctc cagccagata
     2281 ctccccagtt tctaaagcag tacttggaga tgatgaaatt acaagggaac ctagaaaagt
     2341 tgttcttcat cgtggctcaa cgggccttgg tttcaacatt gtaggaggag aagatggaga
     2401 aggaatattt atttccttta tcttagccgg aggacctgct gatctaagtg gagagctcag
     2461 aaaaggagat cgtattatat cggtaaacag tgttgacctc agagctgcta gtcatgagca
     2521 ggcagcagct gcattgaaaa atgctggcca ggctgtcaca attgttgcac aatatcgacc
     2581 tgaagaatac agtcgttttg aagctaaaat acatgattta cgggagcaga tgatgaatag
     2641 tagtattagt tcagggtcag gttctcttcg aactagccag aagcgatccc tctatgtcag
     2701 agcccttttt gattatgaca agactaaaga cagtgggctt cccagtcagg gactgaactt
     2761 caaatttgga gatatcctcc atgttattaa tgcttctgat gatgaatggt ggcaagccag
     2821 gcaggttaca ccagatggtg agagcgatga ggtcggagtg attcccagta aacgcagagt
     2881 tgagaagaaa gaacgagccc gattaaaaac agtgaaattc aattctaaaa cgagagataa
     2941 aggggagatc cctgacgaca tgggatcaaa aggcctgaag catgtaactt ctaatgccag
     3001 cgatagtgaa agtagttacc gtggtcaaga agaatacgtc ttatcttatg aaccagtgaa
     3061 tcaacaagaa gttaattata ctcgaccagt gatcatattg ggacctatga aagacaggat
     3121 aaatgatgac ttgatctcag aatttcctga caaatttgga tcctgtgttc ctcatacaac
     3181 tagaccaaaa cgagattatg aggtagatgg aagagattat cattttgtga cttcaagaga
     3241 gcagatggaa aaagatatcc aggaacataa attcattgaa gctggccagt ataacaatca
     3301 tctatatgga acaagtgttc agtctgtacg agaagtagca gaaaagggca aacactgtat
     3361 ccttgatgtg tctggaaatg ccataaagag attacagatt gcacagcttt accctatctc
     3421 catttttatt aaacccaaat ccatggaaaa tatcatggaa atgaataagc gtctaacaga
     3481 agaacaagcc agaaaaacat ttgagagagc catgaaactg gaacaggagt ttactgaaca
     3541 tttcacagct attgtacagg gggatacgct ggaagacatt tacaaccaag tgaaacagat
     3601 catagaagaa caatctggtt cttacatctg ggttccggca aaagaaaagc tactcgagga
     3661 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga
     3721 caagtagtga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     3781 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3841 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3901 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3961 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     4021 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4081 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4141 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4201 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4261 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4321 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4381 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4441 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4501 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4561 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4621 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4681 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4741 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4801 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4861 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4921 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4981 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5041 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5101 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5161 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5221 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5281 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5341 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5401 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5461 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5521 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5581 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5641 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5701 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5761 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5821 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5881 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5941 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     6001 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6061 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6121 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6181 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6241 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6301 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6361 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6421 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6481 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6541 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6601 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6661 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6721 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6781 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6841 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6901 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     6961 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7021 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7081 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7141 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7201 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7261 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7321 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7381 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7441 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7501 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7561 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7621 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7681 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7741 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7801 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7861 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7921 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7981 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     8041 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     8101 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     8161 tc
//
