LOCUS       sequence-292525-        6479 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged human Src-S4-KD in mammalian cells for 
            structure-function studies.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6479)
  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 6479)
  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-KD
FEATURES             Location/Qualifiers
     source          1..6479
                     /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(1860..1884)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1895..1921
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1925..1951
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1977..2018
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2028..2045
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2068..2085)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2074..2298
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2344..2772
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2431..2450)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2641..2662
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2781..2801)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2786..3115
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2966..3101
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3028..3047
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3182..3976
                     /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(3236..3255)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3846..3865
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4152..4273
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4189..4208)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4243..4262
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4322..4338)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4322..4338)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4335..4357)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4346..4362
                     /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(4370..4376,4377..4394,4395..4400))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4415..4436
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4553..4570)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4724..5312)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4804..4823)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5483..6274,6275..6343))
                     /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     6106..6125
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6344..6448)
                     /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 agcaagccgc agactcaggg
     1021 cctggccaag gatgcctggg agatccctcg ggagtcgctg cggctggagg tcaagctggg
     1081 ccagggctgc tttggcgagg tgtggatggg gacctggaac ggtaccacca gggtggccat
     1141 caaaaccctg aagcctggca cgatgtctcc agaggccttc ctgcaggagg cccaggtcat
     1201 gaagaagctg aggcatgaga agctggtgca gttgtatgct gtggtttcag aggagcccat
     1261 ttacatcgtc acggagtaca tgagcaaggg gagtttgctg gactttctca agggggagac
     1321 aggcaagtac ctgcggctgc ctcagctggt ggacatggct gctcagatcg cctcaggcat
     1381 ggcgtacgtg gagcggatga actacgtcca ccgggacctt cgtgcagcca acatcctggt
     1441 gggagagaac ctggtgtgca aagtggccga ctttgggctg gctcggctca ttgaagacaa
     1501 tgagtacacg gcgcggcaag gtgccaaatt ccccatcaag tggacggctc cagaagctgc
     1561 cctctatggc cgcttcacca tcaagtcgga cgtgtggtcc ttcgggatcc tgctgactga
     1621 gctcaccaca aagggacggg tgccctaccc tgggatggtg aaccgcgagg tgctggacca
     1681 ggtggagcgg ggctaccgga tgccctgccc gccggagtgt cccgagtccc tgcacgacct
     1741 catgtgccag tgctggcgga aggagcctga ggagcggccc accttcgagt acctgcaggc
     1801 cttcctggag gactacttca cgtccaccga gccccagtac cagcccgggg agaacctcga
     1861 cccagctttc ttgtacaaag tggttgatct agagtaccca tacgatgttc cagattacgc
     1921 tggataccca tacgatgttc cagattacgc ttgatctaga gggcccgcgg ttcgaaggta
     1981 agcctatccc taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc
     2041 accattgagt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     2101 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     2161 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     2221 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     2281 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     2341 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2401 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2461 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2521 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     2581 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2641 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     2701 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     2761 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     2821 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2881 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2941 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3001 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     3061 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3121 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     3181 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3241 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3301 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3361 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3421 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3481 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3541 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3601 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3661 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     3721 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3781 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3841 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3901 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3961 tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca
     4021 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4081 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4141 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4201 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4261 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     4321 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4381 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4441 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4501 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4561 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4621 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4681 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4741 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4801 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4861 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4921 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4981 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5041 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5101 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5161 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5221 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5281 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5341 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5401 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5461 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5521 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5581 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5641 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5701 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5761 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5821 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5881 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5941 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6001 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6061 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6121 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6181 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6241 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6301 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6361 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6421 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
