LOCUS       sequence-292527-        6326 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged human Src-S4-UD-S32 in mammalian cells for 
            structure-function studies.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6326)
  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 6326)
  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-UD-S32
FEATURES             Location/Qualifiers
     source          1..6326
                     /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(1707..1731)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1742..1768
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1772..1798
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1824..1865
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1875..1892
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1915..1932)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1921..2145
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2191..2619
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2278..2297)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2488..2509
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2628..2648)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2633..2962
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2813..2948
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2875..2894
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3029..3823
                     /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(3083..3102)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3693..3712
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3999..4120
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4036..4055)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4090..4109
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4169..4185)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4169..4185)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4182..4204)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4193..4209
                     /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(4217..4223,4224..4241,4242..4247))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4262..4283
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4400..4417)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4571..5159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4651..4670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5330..6121,6122..6190))
                     /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     5953..5972
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6191..6295)
                     /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 cgtccgaccc agctttcttg tacaaagtgg ttgatctaga
     1741 gtacccatac gatgttccag attacgctgg atacccatac gatgttccag attacgcttg
     1801 atctagaggg cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt
     1861 ctacgcgtac cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga
     1921 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     1981 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2041 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2101 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     2161 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     2221 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2281 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2341 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2401 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2461 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     2521 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     2581 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     2641 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2701 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2761 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2821 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     2881 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     2941 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     3001 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3061 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3121 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3181 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3241 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3301 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3361 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3421 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3481 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3541 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     3601 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     3661 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     3721 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     3781 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     3841 gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     3901 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     3961 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4021 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4081 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4141 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4201 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4261 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4321 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4381 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4441 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4501 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4561 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4621 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4681 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4741 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4801 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4861 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4921 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4981 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5041 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5101 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5161 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5221 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5281 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5341 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5401 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5461 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5521 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5581 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5641 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5701 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5761 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5821 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5881 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5941 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6001 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6061 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6121 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6181 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6241 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6301 ttccccgaaa agtgccacct gacgtc
//
