LOCUS       sequence-292526-        7013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged human Src-S4-S32-KD in mammalian cells for 
            structure-function studies.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7013)
  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 7013)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2018.09"; journalName: "Cell Rep"; date: 
            "2018-10-9- 9"; volume: "25"; issue: "2"; pages: "449-463.e4"
COMMENT     Sequence Label: pDEST40-2XHA-Src-S4-S32-KD
FEATURES             Location/Qualifiers
     source          1..7013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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(2394..2418)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2429..2455
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2459..2485
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2511..2552
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2562..2579
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2602..2619)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2608..2832
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2878..3306
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2965..2984)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3175..3196
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3315..3335)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3320..3649
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3500..3635
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3562..3581
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3716..4510
                     /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(3770..3789)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4380..4399
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4686..4807
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4723..4742)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4777..4796
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4856..4872)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4856..4872)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4869..4891)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4880..4896
                     /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(join(4904..4910,4911..4928,4929..4934))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4949..4970
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5087..5104)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5258..5846)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5338..5357)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6017..6808,6809..6877))
                     /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     6640..6659
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6878..6982)
                     /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 agcacctccc cgcagagggc
     1021 gggcccgctg gccggtggag tgaccacctt tgtggccctc tatgactatg agtctaggac
     1081 ggagacagac ctgtccttca agaaaggcga gcggctccag attgtcaaca acacagaggg
     1141 agactggtgg ctggcccact cgctcagcac aggacagaca ggctacatcc ccagcaacta
     1201 cgtggcgccc tccgactcca tccaggctga ggagtggtat tttggcaaga tcaccagacg
     1261 ggagtcagag cggttactgc tcaatgcaga gaacccgaga gggaccttcc tcgtgcgaga
     1321 aagtgagacc acgaaaggtg cctactgcct ctcagtgtct gacttcgaca acgccaaggg
     1381 cctcaacgtg aagcactaca agatccgcaa gctggacagc ggcggcttct acatcacctc
     1441 ccgcacccag ttcaacagcc tgcagcagct ggtggcctac tactccaaac acgccgatgg
     1501 cctgtgccac cgcctcacca ccgtgtgccc cacgtccaag ccgcagactc agggcctggc
     1561 caaggatgcc tgggagatcc ctcgggagtc gctgcggctg gaggtcaagc tgggccaggg
     1621 ctgctttggc gaggtgtgga tggggacctg gaacggtacc accagggtgg ccatcaaaac
     1681 cctgaagcct ggcacgatgt ctccagaggc cttcctgcag gaggcccagg tcatgaagaa
     1741 gctgaggcat gagaagctgg tgcagttgta tgctgtggtt tcagaggagc ccatttacat
     1801 cgtcacggag tacatgagca aggggagttt gctggacttt ctcaaggggg agacaggcaa
     1861 gtacctgcgg ctgcctcagc tggtggacat ggctgctcag atcgcctcag gcatggcgta
     1921 cgtggagcgg atgaactacg tccaccggga ccttcgtgca gccaacatcc tggtgggaga
     1981 gaacctggtg tgcaaagtgg ccgactttgg gctggctcgg ctcattgaag acaatgagta
     2041 cacggcgcgg caaggtgcca aattccccat caagtggacg gctccagaag ctgccctcta
     2101 tggccgcttc accatcaagt cggacgtgtg gtccttcggg atcctgctga ctgagctcac
     2161 cacaaaggga cgggtgccct accctgggat ggtgaaccgc gaggtgctgg accaggtgga
     2221 gcggggctac cggatgccct gcccgccgga gtgtcccgag tccctgcacg acctcatgtg
     2281 ccagtgctgg cggaaggagc ctgaggagcg gcccaccttc gagtacctgc aggccttcct
     2341 ggaggactac ttcacgtcca ccgagcccca gtaccagccc ggggagaacc tcgacccagc
     2401 tttcttgtac aaagtggttg atctagagta cccatacgat gttccagatt acgctggata
     2461 cccatacgat gttccagatt acgcttgatc tagagggccc gcggttcgaa ggtaagccta
     2521 tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg tcatcatcac catcaccatt
     2581 gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     2641 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2701 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2761 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     2821 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     2881 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2941 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3001 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3061 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3121 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3181 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3241 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3301 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3361 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3421 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3481 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3541 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     3601 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     3661 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     3721 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3781 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3841 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     3901 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3961 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4021 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4081 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4141 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4201 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     4261 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4321 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4381 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4441 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4501 gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc
     4561 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4621 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4681 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4741 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4801 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     4861 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4921 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4981 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5041 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5101 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5161 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5221 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5281 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5341 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5401 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5461 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5521 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5581 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5641 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5701 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5761 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5821 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5881 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5941 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6001 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6061 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6121 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6181 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6241 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6301 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6361 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6421 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6481 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6541 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6601 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6661 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6721 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6781 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6841 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6901 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6961 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
