LOCUS       sequence-478757-        6207 bp    DNA     circular SYN 16-JUL-2025
DEFINITION  S.cerevisiae Sla1-SH3#1-3 (5-413) in pGEX6P1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6207)
  TITLE     Competitive Binding of Actin and SH3 domains at Proline-rich regions
            of Las17/WASP Regulates Actin Polymerisation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6207)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: PKA1248
FEATURES             Location/Qualifiers
     source          1..6207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(184..189,190..205,206..212)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    220..236
                     /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)."
     CDS             259..912
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     870..892
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             919..942
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1720..1731
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2258..2280)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2425..2443)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2511..2615
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2616..2684,2685..3476)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2834..2853)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3647..4235
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4136..4155
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4389..4406
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4479..4556
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4557..5639
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4576..4595)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5688..5693,5694..5711,5712..5718)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5726..5742
                     /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)."
     primer_bind     5750..5766
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5750..5766
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5762..6207,1..82)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5782..5799)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5782..5798)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5791..5813)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tagcttatcg actgcacggt gcaccaatgc ttctggcgtc aggcagccat cggaagctgt
       61 ggtatggctg tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg cgcactcccg
      121 ttctggataa tgttttttgc gccgacatca taacggttct ggcaaatatt ctgaaatgag
      181 ctgttgacaa ttaatcatcg gctcgtataa tgtgtggaat tgtgagcgga taacaatttc
      241 acacaggaaa cagtattcat gtcccctata ctaggttatt ggaaaattaa gggccttgtg
      301 caacccactc gacttctttt ggaatatctt gaagaaaaat atgaagagca tttgtatgag
      361 cgcgatgaag gtgataaatg gcgaaacaaa aagtttgaat tgggtttgga gtttcccaat
      421 cttccttatt atattgatgg tgatgttaaa ttaacacagt ctatggccat catacgttat
      481 atagctgaca agcacaacat gttgggtggt tgtccaaaag agcgtgcaga gatttcaatg
      541 cttgaaggag cggttttgga tattagatac ggtgtttcga gaattgcata tagtaaagac
      601 tttgaaactc tcaaagttga ttttcttagc aagctacctg aaatgctgaa aatgttcgaa
      661 gatcgtttat gtcataaaac atatttaaat ggtgatcatg taacccatcc tgacttcatg
      721 ttgtatgacg ctcttgatgt tgttttatac atggacccaa tgtgcctgga tgcgttccca
      781 aaattagttt gttttaaaaa acgtattgaa gctatcccac aaattgataa gtacttgaaa
      841 tccagcaagt atatagcatg gcctttgcag ggctggcaag ccacgtttgg tggtggcgac
      901 catcctccaa aatcggatct ggaagttctg ttccaggggc ccctgggatc cctgggcatc
      961 tatagggccg tctatgccta tgagccgcag acaccagaag aactggccat ccaagaagac
     1021 gatctgttgt acctcttaca gaagtcagac attgacgatt ggtggacagt aaagaaaaga
     1081 gtcattggtt ccgatagcga agaaccggtg ggtctagtgc cctccactta cattgaagaa
     1141 gctcctgttt tgaagaaggt aagagccatt tatgattatg aacaggtgca aaatgctgat
     1201 gaagaattga cgtttcatga aaatgacgtt tttgatgtat tcgatgataa agatgctgat
     1261 tggctgttgg ttaagtctac cgtttccaat gaattcggct tcattccagg caattacgtc
     1321 gaaccagaga atgggtccac ttccaagcag gaacaggctc ccgctgccgc tgaagctcct
     1381 gcagctactc ctgcagccgc tcctgcttct gctgccgtac tacccaccaa ctttttacca
     1441 cctcctcaac acaatgatag agctcgtatg atgcaaagta aagaagacca agctccagat
     1501 gaggatgaag agggccctcc tcctgccatg cctgcaagac caacggctac taccgaaact
     1561 actgatgcca ccgctgcggc tgttcgatct cgtacaagac tatcatacag cgataacgat
     1621 aatgacgatg aagaggatga ttactattac aacagcaaca gcaacaacgt tggaaatcac
     1681 gaatacaata cagagtatca ctcttggaac gttaccgaaa tcgagggaag gaaaaagaag
     1741 aaggccaagc tatcaatcgg taacaacaag ataaacttca taccccaaaa ggggactcct
     1801 catgagtggt caattgataa attggtctct tatgataacg aaaagaaaca catgttcttg
     1861 gaatttgttg atccatatag aagccttgaa ttacacactg gtaataccac cacatgtgaa
     1921 gagatcatga acattattgg tgaatataaa ggtgcttccc gtgatcctgg gttgagagaa
     1981 gtcgaaatgg cttccaaatc caaaaagaga ggtattgttc aatatgactt catggctgaa
     2041 tcacaggacg aattaaccat aaaatcaggc gataaagtct acattttaga tgataaaaaa
     2101 tctaaggact ggtggatgtg ccaactggtt gattcaggga aaagcggtct cgttcctgca
     2161 cagtttattg agcctgttcg ttgatgagtc gactcgagcg gccgcatcgt gactgactga
     2221 cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     2281 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     2341 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
     2401 tgtataattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     2461 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2521 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2581 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     2641 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2701 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     2761 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     2821 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     2881 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     2941 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3001 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3061 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3121 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
     3181 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3241 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3301 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3361 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3421 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3481 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3541 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3601 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3661 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3721 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3781 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3841 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3901 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3961 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4021 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4081 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4141 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4201 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4261 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4321 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4381 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     4441 ttacgcatct gtgcggtatt tcacaccgca taaattccga caccatcgaa tggcgcaaaa
     4501 cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga
     4561 aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc
     4621 gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga
     4681 tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt
     4741 tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg
     4801 cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa
     4861 gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc
     4921 tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta
     4981 atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt attattttct
     5041 cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa
     5101 tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc
     5161 ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg
     5221 ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga
     5281 tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc
     5341 tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt
     5401 atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg
     5461 accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct
     5521 cactggtgaa aagaaaaacc acgctagcgc ccaatacgca aaccgcctct ccccgcgcgt
     5581 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     5641 cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cccaggcttt acactttatg
     5701 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     5761 tatgaccatg attacggatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     5821 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     5881 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     5941 ctttgcctgg tttccggcac cagaagcggt gccggaaagc tggctggagt gcgatcttcc
     6001 tgaggccgat actgtcgtcg tcccctcaaa ctggcagatg cacggttacg atgcgcccat
     6061 ctacaccaac gtgacctatc ccattacggt caatccgccg tttgttccca cggagaatcc
     6121 gacgggttgt tactcgctca catttaatgt tgatgaaagc tggctacagg aaggccagac
     6181 gcgaattatt tttgatggcg ttggaat
//
