LOCUS       sequence-243616-        7502 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human c-Src with Y530F mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7502)
  AUTHORS   Zhang Y, Cao F, Zhou Y, Feng Z, Sit B, Krendel M, Yu CH
  TITLE     Tail domains of myosin-1e regulate phosphatidylinositol signaling 
            and F-actin polymerization at the ventral layer of podosomes.
  JOURNAL   Mol Biol Cell. 2019 Mar 1;30(5):622-635. doi: 
            10.1091/mbc.E18-06-0398. Epub 2019 Jan 2.
  PUBMED    30601698
REFERENCE   2  (bases 1 to 7502)
  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: 
            "10.1091/mbc.E18-06-0398"; journalName: "Mol Biol Cell"; date: 
            "2019-03-1- 1"; volume: "30"; issue: "5"; pages: "622-635"
COMMENT     Sequence Label: Src-Y530F
FEATURES             Location/Qualifiers
     source          1..7502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        123..1301
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          354..1292
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1249..1269
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1318..1337
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1318..1336
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3156..3185
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3201..3218
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3241..3258)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3247..3471
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3517..3945
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3604..3623)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3814..3835
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3954..3974)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3959..4288
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4139..4274
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4201..4220
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4336..4383
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4402..4800
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4958..5079
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4995..5014)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5049..5068
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5128..5144)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5128..5144)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5141..5163)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5152..5168
                     /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(5176..5182,5183..5200,5201..5206))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5221..5242
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5359..5376)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5519..6107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5599..5618)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6278..7069,7070..7138))
                     /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     6901..6920
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7139..7243)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7318..7337)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgcgttgaca ttgattattg actaggcttt tgcaaaaagc tttgcaaaga tggataaagt
       61 tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa
      121 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
      181 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
      241 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
      301 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
      361 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
      421 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
      481 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
      541 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
      601 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
      661 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
      721 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
      781 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
      841 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
      901 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
      961 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     1021 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     1081 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     1141 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     1201 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     1261 cagtggttca aagttttttt cttccatttc aggtgtcgtg aggaattagc ttggtactaa
     1321 tacgactcac tatagggaga cccaagctgg ctaggtaagc ttggtaccga gctcggatcc
     1381 agtgtggtgg aattcgatca tgggtagcaa caagagcaag cccaaggatg ccagccagcg
     1441 gcgccgcagc ctggagcccg ccgagaacgt gcacggcgct ggcgggggcg ctttccccgc
     1501 ctcgcagacc cccagcaagc cagcctcggc cgacggccac cgcggcccca gcgcggcctt
     1561 cgcccccgcg gccgccgagc ccaagctgtt cggaggcttc aactcctcgg acaccgtcac
     1621 ctccccgcag agggcgggcc cgctggccgg tggagtgacc acctttgtgg ccctctatga
     1681 ctatgagtct aggacggaga cagacctgtc cttcaagaaa ggcgagcggc tccagattgt
     1741 caacaacaca gagggagact ggtggctggc ccactcgctc agcacaggac agacaggcta
     1801 catccccagc aactacgtgg cgccctccga ctccatccag gctgaggagt ggtattttgg
     1861 caagatcacc agacgggagt cagagcggtt actgctcaat gcagagaacc cgagagggac
     1921 cttcctcgtg cgagaaagtg agaccacgaa aggtgcctac tgcctctcag tgtctgactt
     1981 cgacaacgcc aagggcctca acgtgaagca ctacaagatc cgcaagctgg acagcggcgg
     2041 cttctacatc acctcccgca cccagttcaa cagcctgcag cagctggtgg cctactactc
     2101 caaacacgcc gatggcctgt gccaccgcct caccaccgtg tgccccacgt ccaagccgca
     2161 gactcagggc ctggccaagg atgcctggga gatccctcgg gagtcgctgc ggctggaggt
     2221 caagctgggc cagggctgct ttggcgaggt gtggatgggg acctggaacg gtaccaccag
     2281 ggtggccatc aaaaccctga agcctggcac gatgtctcca gaggccttcc tgcaggaggc
     2341 ccaggtcatg aagaagctga ggcatgagaa gctggtgcag ttgtatgctg tggtttcaga
     2401 ggagcccatt tacatcgtca cggagtacat gagcaagggg agtttgctgg actttctcaa
     2461 gggggagaca ggcaagtacc tgcggctgcc tcagctggtg gacatggctg ctcagatcgc
     2521 ctcaggcatg gcgtacgtgg agcggatgaa ctacgtccac cgggaccttc gtgcagccaa
     2581 catcctggtg ggagagaacc tggtgtgcaa agtggccgac tttgggctgg ctcggctcat
     2641 tgaagacaat gagtacacgg cgcggcaagg tgccaaattc cccatcaagt ggacggctcc
     2701 agaagctgcc ctctatggcc gcttcaccat caagtcggac gtgtggtcct tcgggatcct
     2761 gctgactgag ctcaccacaa agggacgggt gccctaccct gggatggtga accgcgaggt
     2821 gctggaccag gtggagcggg gctaccggat gccctgcccg ccggagtgtc ccgagtccct
     2881 gcacgacctc atgtgccagt gctggcggaa ggagcctgag gagcggccca ccttcgagta
     2941 cctgcaggcc ttcctggagg actacttcac gtccaccgag ccccagttcc agcctgggga
     3001 gaacctctag gcacaggcgg gcccagaccg gcttctcggc ttggatcgat cctgctccct
     3061 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa ttctgcagat atccagcaca
     3121 gtggcggccg ctcgagtcta gagggcccgc ggttcgaaca aaaactcatc tcagaagagg
     3181 atctgaatat gcataccggt catcatcacc atcaccattg agtttaaacc cgctgatcag
     3241 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     3301 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     3361 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     3421 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     3481 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     3541 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3601 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3661 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3721 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3781 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3841 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     3901 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     3961 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4021 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4081 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4141 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4201 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     4261 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     4321 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac
     4381 gacaaggtga ggaactaaac catggccaag cctttgtctc aagaagaatc caccctcatt
     4441 gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag cgtcgccagc
     4501 gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca ttttactggg
     4561 ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc tggcaacctg
     4621 acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg cggacggtgc
     4681 cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga cagtgatgga
     4741 cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa
     4801 gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc
     4861 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4921 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4981 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5041 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5101 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5161 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5221 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5281 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5341 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5401 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5461 aggaaagaac atgtgagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5521 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5581 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5641 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5701 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5761 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5821 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5881 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5941 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6001 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6061 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6121 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6181 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6241 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6301 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6361 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6421 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6481 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6541 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6601 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6661 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6721 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6781 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6841 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6901 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6961 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7021 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7081 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7141 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7201 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7261 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     7321 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     7381 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     7441 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     7501 ta
//
