LOCUS       sequence-363993-        7007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Flag-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 7007)
  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 7007)
  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: pDEST40-2XFL-Src-S4-S32-KD
FEATURES             Location/Qualifiers
     source          1..7007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        146..525
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        526..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     774..793
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        774..792
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    822..846
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2305..2329)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2340..2363
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2367..2390
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2416..2457
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2467..2484
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2507..2524)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2513..2737
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2783..3211
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2870..2889)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3080..3101
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3220..3240)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3225..3554
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3405..3540
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3467..3486
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3621..4415
                     /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(3675..3694)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4285..4304
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4591..4712
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4628..4647)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4682..4701
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4761..4777)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4761..4777)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4774..4796)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4785..4801
                     /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(4809..4815,4816..4833,4834..4839))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4854..4875
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4992..5009)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5163..5751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5243..5262)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5922..6713,6714..6782))
                     /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     6545..6564
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6783..6887)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6962..6981)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
       61 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
      121 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
      181 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      241 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      301 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      361 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      421 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      481 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      541 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      601 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      661 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg
      781 actcactata gggagaccca agctggctag ttaagctatc aacaagtttg tacaaaaaag
      841 caggcttcga aggagataga accatgggta gcaacaagag caagcccaag gatgccagcc
      901 agcggcgccg cagcacctcc ccgcagaggg cgggcccgct ggccggtgga gtgaccacct
      961 ttgtggccct ctatgactat gagtctagga cggagacaga cctgtccttc aagaaaggcg
     1021 agcggctcca gattgtcaac aacacagagg gagactggtg gctggcccac tcgctcagca
     1081 caggacagac aggctacatc cccagcaact acgtggcgcc ctccgactcc atccaggctg
     1141 aggagtggta ttttggcaag atcaccagac gggagtcaga gcggttactg ctcaatgcag
     1201 agaacccgag agggaccttc ctcgtgcgag aaagtgagac cacgaaaggt gcctactgcc
     1261 tctcagtgtc tgacttcgac aacgccaagg gcctcaacgt gaagcactac aagatccgca
     1321 agctggacag cggcggcttc tacatcacct cccgcaccca gttcaacagc ctgcagcagc
     1381 tggtggccta ctactccaaa cacgccgatg gcctgtgcca ccgcctcacc accgtgtgcc
     1441 ccacgtccaa gccgcagact cagggcctgg ccaaggatgc ctgggagatc cctcgggagt
     1501 cgctgcggct ggaggtcaag ctgggccagg gctgctttgg cgaggtgtgg atggggacct
     1561 ggaacggtac caccagggtg gccatcaaaa ccctgaagcc tggcacgatg tctccagagg
     1621 ccttcctgca ggaggcccag gtcatgaaga agctgaggca tgagaagctg gtgcagttgt
     1681 atgctgtggt ttcagaggag cccatttaca tcgtcacgga gtacatgagc aaggggagtt
     1741 tgctggactt tctcaagggg gagacaggca agtacctgcg gctgcctcag ctggtggaca
     1801 tggctgctca gatcgcctca ggcatggcgt acgtggagcg gatgaactac gtccaccggg
     1861 accttcgtgc agccaacatc ctggtgggag agaacctggt gtgcaaagtg gccgactttg
     1921 ggctggctcg gctcattgaa gacaatgagt acacggcgcg gcaaggtgcc aaattcccca
     1981 tcaagtggac ggctccagaa gctgccctct atggccgctt caccatcaag tcggacgtgt
     2041 ggtccttcgg gatcctgctg actgagctca ccacaaaggg acgggtgccc taccctggga
     2101 tggtgaaccg cgaggtgctg gaccaggtgg agcggggcta ccggatgccc tgcccgccgg
     2161 agtgtcccga gtccctgcac gacctcatgt gccagtgctg gcggaaggag cctgaggagc
     2221 ggcccacctt cgagtacctg caggccttcc tggaggacta cttcacgtcc accgagcccc
     2281 agtaccagcc cggggagaac ctcgacccag ctttcttgta caaagtggtt gatctagagg
     2341 attacaagga tgacgacgat aagggagatt acaaggatga cgacgataag tgatctagag
     2401 ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc tcggtctcga ttctacgcgt
     2461 accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct
     2521 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2581 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2641 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2701 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2761 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2821 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2881 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2941 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3001 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3061 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3121 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3181 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3241 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3301 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3361 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3421 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3481 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3541 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3601 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3661 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3721 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3781 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3841 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3901 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3961 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4021 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4081 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4141 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4201 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4261 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4321 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4381 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg
     4441 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4501 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4561 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     4621 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4681 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     4741 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4801 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4861 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4921 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4981 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5041 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5101 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5161 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5221 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5281 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5341 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5401 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5461 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5521 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5581 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5641 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5701 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5761 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5821 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5881 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5941 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6001 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6061 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6121 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6181 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6241 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6301 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6361 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6421 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6481 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6541 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6601 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6661 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6721 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6781 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6841 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6901 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
     6961 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccct
//
