LOCUS       Exported                7181 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses HA-tagged human Src-K298M-Y419F-Y530F in mammalian cells 
            for structure-function studies.
ACCESSION   .
VERSION     .
KEYWORDS    pDEST40-2XHA-Src-K298M-Y419F-Y530F
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7181)
  AUTHORS   Spassov DS, Ruiz-Saenz A, Piple A, Moasser MM
  TITLE     A Dimerization Function in the Intrinsically Disordered N-Terminal 
            Region of Src.
  JOURNAL   Cell Rep. 2018 Oct 9;25(2):449-463.e4. doi: 
            10.1016/j.celrep.2018.09.035.
  PUBMED    30304684
REFERENCE   2  (bases 1 to 7181)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7181
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     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"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    911..935
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2562..2586)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2597..2623
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2627..2653
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2679..2720
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2730..2747
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2770..2787)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2776..3000
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3046..3474
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3133..3152)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3343..3364
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3483..3503)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3488..3817
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3668..3803
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3730..3749
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3884..4678
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3938..3957)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4548..4567
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4854..4975
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4891..4910)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4945..4964
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5024..5040)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5024..5040)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5037..5059)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5048..5064
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5072..5102)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5117..5138
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5255..5272)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5426..6014)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5506..5525)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6185..7045)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6808..6827
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7046..7150)
                     /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 gctggctagt
      901 taagctatca acaagtttgt acaaaaaagc aggcttcgaa ggagatagaa ccatgggtag
      961 caacaagagc aagcccaagg atgccagcca gcggcgccgc agcctggagc ccgccgagaa
     1021 cgtgcacggc gctggcgggg gcgctttccc cgcctcgcag acccccagca agccagcctc
     1081 ggccgacggc caccgcggcc ccagcgcggc cttcgccccc gcggccgccg agcccaagct
     1141 gttcggaggc ttcaactcct cggacaccgt cacctccccg cagagggcgg gcccgctggc
     1201 cggtggagtg accacctttg tggccctcta tgactatgag tctaggacgg agacagacct
     1261 gtccttcaag aaaggcgagc ggctccagat tgtcaacaac acagagggag actggtggct
     1321 ggcccactcg ctcagcacag gacagacagg ctacatcccc agcaactacg tggcgccctc
     1381 cgactccatc caggctgagg agtggtattt tggcaagatc accagacggg agtcagagcg
     1441 gttactgctc aatgcagaga acccgagagg gaccttcctc gtgcgagaaa gtgagaccac
     1501 gaaaggtgcc tactgcctct cagtgtctga cttcgacaac gccaagggcc tcaacgtgaa
     1561 gcactacaag atccgcaagc tggacagcgg cggcttctac atcacctccc gcacccagtt
     1621 caacagcctg cagcagctgg tggcctacta ctccaaacac gccgatggcc tgtgccaccg
     1681 cctcaccacc gtgtgcccca cgtccaagcc gcagactcag ggcctggcca aggatgcctg
     1741 ggagatccct cgggagtcgc tgcggctgga ggtcaagctg ggccagggct gctttggcga
     1801 ggtgtggatg gggacctgga acggtaccac cagggtggcc atcatgaccc tgaagcctgg
     1861 cacgatgtct ccagaggcct tcctgcagga ggcccaggtc atgaagaagc tgaggcatga
     1921 gaagctggtg cagttgtatg ctgtggtttc agaggagccc atttacatcg tcacggagta
     1981 catgagcaag gggagtttgc tggactttct caagggggag acaggcaagt acctgcggct
     2041 gcctcagctg gtggacatgg ctgctcagat cgcctcaggc atggcgtacg tggagcggat
     2101 gaactacgtc caccgggacc ttcgtgcagc caacatcctg gtgggagaga acctggtgtg
     2161 caaagtggcc gactttgggc tggctcggct cattgaagac aatgagttca cggcgcggca
     2221 aggtgccaaa ttccccatca agtggacggc tccagaagct gccctctatg gccgcttcac
     2281 catcaagtcg gacgtgtggt ccttcgggat cctgctgact gagctcacca caaagggacg
     2341 ggtgccctac cctgggatgg tgaaccgcga ggtgctggac caggtggagc ggggctaccg
     2401 gatgccctgc ccgccggagt gtcccgagtc cctgcacgac ctcatgtgcc agtgctggcg
     2461 gaaggagcct gaggagcggc ccaccttcga gtacctgcag gccttcctgg aggactactt
     2521 cacgtccacc gagccccagt tccagcccgg ggagaacctc gacccagctt tcttgtacaa
     2581 agtggttgat ctagagtacc catacgatgt tccagattac gctggatacc catacgatgt
     2641 tccagattac gcttgatcta gagggcccgc ggttcgaagg taagcctatc cctaaccctc
     2701 tcctcggtct cgattctacg cgtaccggtc atcatcacca tcaccattga gtttaaaccc
     2761 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2821 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2881 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     2941 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     3001 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     3061 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3121 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3181 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3241 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     3301 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3361 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     3421 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     3481 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3541 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3601 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3661 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3721 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     3781 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     3841 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     3901 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     3961 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4021 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     4081 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4141 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4201 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4261 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4321 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4381 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     4441 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4501 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4561 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4621 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4681 gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4741 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4801 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4861 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4921 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4981 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5041 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5101 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5161 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5221 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5281 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5341 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5401 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5461 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5521 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5581 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5641 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5701 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5761 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5821 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5881 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5941 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6001 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6061 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6121 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6181 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6241 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6301 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6361 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6421 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6481 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6541 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6601 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6661 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6721 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6781 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6841 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6901 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6961 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7021 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7081 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7141 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
