LOCUS       40924_14270        7173 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian vector pdcDNA-HA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7173)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7173)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 7173)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7173)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 
            9052, BELGIUM"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             957..983
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     protein_bind    987..1111
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1136..1166
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1220..1876
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2221..2523
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2567..2691)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2726..2744)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2770..2994
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3040..3468
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3482..3811
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3878..4669
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4846..4979
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5016..5032)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5040..5056)
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5064..5094)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5109..5130)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5418..6006)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6180..7037)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7038..7142)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttctgcagat atcatgtacc
      961 catacgacgt cccagactac gctatcacaa gtttgtacaa aaaagctgaa cgagaaacgt
     1021 aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag actacataat
     1081 actgtaaaac acaacatatc cagtcactat ggcggccgca ttaggcaccc caggctttac
     1141 actttatgct tccggctcgt ataatgtgtg gattttgagt taggatccgg cgagattttc
     1201 aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc
     1261 ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa
     1321 ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa
     1381 gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg
     1441 tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt
     1501 tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg
     1561 gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt
     1621 ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac
     1681 cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg
     1741 caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc
     1801 cgtctgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga
     1861 gtggcagggc ggggcgtaaa cgcgtggatc cggcttacta aaagccagat aacagtatgc
     1921 gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat
     1981 gtcaaaaaga ggtgtgctat gaagcagcgt attacagtga cagttgacag cgacagctat
     2041 cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga
     2101 atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg
     2161 aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg gactggtgaa
     2221 atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta
     2281 cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt
     2341 ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc
     2401 tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg
     2461 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga
     2521 atataaatgt caggctccct tatacacagc cagtctgcag gtcgaccata gtgactggat
     2581 atgttgtgtt ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt
     2641 gatatttata tcattttacg tttctcgttc agctttcttg tacaaagtgg tgattcccct
     2701 cgagcatgca tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat
     2761 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2821 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2881 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2941 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3001 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3061 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3121 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3181 aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3241 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3301 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3361 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3421 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     3481 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3541 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3601 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3661 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3721 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     3781 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     3841 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3901 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3961 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4021 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4081 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4141 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4201 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4261 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4321 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4381 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4441 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4501 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4561 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4621 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4681 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4741 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4801 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4861 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4921 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4981 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5041 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5101 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5161 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5221 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5281 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5341 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5401 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5461 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5521 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5581 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     5641 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5701 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5761 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5821 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     5881 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5941 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6001 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6061 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6121 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6181 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6241 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6301 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6361 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6421 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6481 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6541 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6601 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6661 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6721 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6781 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6841 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6901 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6961 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7021 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7081 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7141 cgcacatttc cccgaaaagt gccacctgac gtc
//
