LOCUS       sequence-291890-        6965 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Flag-tagged human Src-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 6965)
  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 6965)
  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-S32-KD
FEATURES             Location/Qualifiers
     source          1..6965
                     /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(2352..2376)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2387..2410
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2414..2437
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2463..2504
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2514..2531
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2554..2571)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2560..2784
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2830..3258
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2917..2936)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3127..3148
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3267..3287)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3272..3601
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3452..3587
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3514..3533
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3668..4462
                     /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(3722..3741)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4332..4351
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4638..4759
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4675..4694)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4729..4748
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4808..4824)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4808..4824)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4821..4843)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4832..4848
                     /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(4856..4862,4863..4880,4881..4886))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4901..4922
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5039..5056)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5210..5798)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5290..5309)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5969..6760,6761..6829))
                     /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     6592..6611
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6830..6934)
                     /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 cacctccccg cagagggcgg gcccgctggc cggtggagtg accacctttg tggccctcta
     1021 tgactatgag tctaggacgg agacagacct gtccttcaag aaaggcgagc ggctccagat
     1081 tgtcaacaac acagagggag actggtggct ggcccactcg ctcagcacag gacagacagg
     1141 ctacatcccc agcaactacg tggcgccctc cgactccatc caggctgagg agtggtattt
     1201 tggcaagatc accagacggg agtcagagcg gttactgctc aatgcagaga acccgagagg
     1261 gaccttcctc gtgcgagaaa gtgagaccac gaaaggtgcc tactgcctct cagtgtctga
     1321 cttcgacaac gccaagggcc tcaacgtgaa gcactacaag atccgcaagc tggacagcgg
     1381 cggcttctac atcacctccc gcacccagtt caacagcctg cagcagctgg tggcctacta
     1441 ctccaaacac gccgatggcc tgtgccaccg cctcaccacc gtgtgcccca cgtccaagcc
     1501 gcagactcag ggcctggcca aggatgcctg ggagatccct cgggagtcgc tgcggctgga
     1561 ggtcaagctg ggccagggct gctttggcga ggtgtggatg gggacctgga acggtaccac
     1621 cagggtggcc atcaaaaccc tgaagcctgg cacgatgtct ccagaggcct tcctgcagga
     1681 ggcccaggtc atgaagaagc tgaggcatga gaagctggtg cagttgtatg ctgtggtttc
     1741 agaggagccc atttacatcg tcacggagta catgagcaag gggagtttgc tggactttct
     1801 caagggggag acaggcaagt acctgcggct gcctcagctg gtggacatgg ctgctcagat
     1861 cgcctcaggc atggcgtacg tggagcggat gaactacgtc caccgggacc ttcgtgcagc
     1921 caacatcctg gtgggagaga acctggtgtg caaagtggcc gactttgggc tggctcggct
     1981 cattgaagac aatgagtaca cggcgcggca aggtgccaaa ttccccatca agtggacggc
     2041 tccagaagct gccctctatg gccgcttcac catcaagtcg gacgtgtggt ccttcgggat
     2101 cctgctgact gagctcacca caaagggacg ggtgccctac cctgggatgg tgaaccgcga
     2161 ggtgctggac caggtggagc ggggctaccg gatgccctgc ccgccggagt gtcccgagtc
     2221 cctgcacgac ctcatgtgcc agtgctggcg gaaggagcct gaggagcggc ccaccttcga
     2281 gtacctgcag gccttcctgg aggactactt cacgtccacc gagccccagt accagcccgg
     2341 ggagaacctc gacccagctt tcttgtacaa agtggttgat ctagaggatt acaaggatga
     2401 cgacgataag ggagattaca aggatgacga cgataagtga tctagagggc ccgcggttcg
     2461 aaggtaagcc tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc
     2521 accatcacca ttgagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     2581 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     2641 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     2701 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     2761 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     2821 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2881 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     2941 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3001 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3061 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3121 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3181 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3241 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3301 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3361 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3421 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3481 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3541 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3601 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     3661 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3721 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3781 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3841 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3901 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3961 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4021 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4081 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4141 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     4201 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4261 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4321 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4381 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4441 ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga
     4501 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4561 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4621 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4681 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4741 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4801 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4861 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4921 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4981 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5041 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5101 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5161 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5221 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5281 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5341 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5401 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5461 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5521 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5581 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5641 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5701 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5761 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5821 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5881 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5941 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6001 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6061 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6121 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6181 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6241 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6301 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6361 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6421 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6481 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6541 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6601 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6661 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6721 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6781 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6841 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6901 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6961 acgtc
//
