LOCUS       sequence-478749-        3225 bp    DNA     circular SYN 16-JUL-2025
DEFINITION  His-Las17 (300-422) pTOPO 6xHis-Las17 (300-422) as a BamH1-HindIII 
            fragment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3225)
  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  JOURNAL   Unpublished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 3225)
  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            "Unpublished"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: PKA1280
FEATURES             Location/Qualifiers
     source          1..3225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(623..1414,1415..1483))
                     /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     1246..1265
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1484..1588)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1614..2069)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1751..1772)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1963..1982
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(2166..2213)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2209..2227
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(join(2661..2675,2676..2684))
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             complement(2688..2720)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             complement(2724..2741)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2761..2783
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2814..2833)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2815..2833)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2918..2935)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(3089..3225,1..452))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3169..3188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
       61 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
      121 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
      181 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
      241 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
      301 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
      361 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
      421 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
      481 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
      541 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
      601 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
      661 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
      721 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
      781 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
      841 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
      901 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
      961 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     1021 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     1081 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     1141 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     1201 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     1261 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     1321 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     1381 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     1441 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     1501 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     1561 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     1621 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     1681 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     1741 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     1801 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     1861 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     1921 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     1981 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     2041 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
     2101 ggaagggcga tcggtgcggg cctcttcgct attacgccag atccggatat agttcctcct
     2161 ttcagcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc tagttattgc
     2221 tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt tgttagcagc cggatcaagc
     2281 ttttagctat ttgtctgata tccaaagggg cggttagaat tattgtactg ttgtggattt
     2341 tgttgcatcg taacgtttgg tgtggggcga gatgctcttg gaggaggtgg tggtgctggc
     2401 cctcttcttc cagtggcctg tggaaggagg ctatttccac cggcagaatt cagggtatta
     2461 ctggttacgt gcctatgtgg tggaggcggt ggtgctggcc cacgccttgg cggaggtgga
     2521 ggagcaggta ggcgaacacc tgaaccttca gtagtggtac gcattggcgg aggaggtggc
     2581 acaggccgat tatttctgtt aggcaactgt ggaaggggcc tattctgctg tggtatgccc
     2641 attgaaggag cttgtggatc cttatcgtca tcgtcgtaca gatcccgacc catttgctgt
     2701 ccaccagtca tgctagccat accatgatga tgatgatgat gagaaccccg catatgtata
     2761 tctccttctt aaagttaaac aaaattattt ctagagggaa accgttgtgg tctccctata
     2821 gtgagtcgta ttaatttcgc gggatcgaga tcctgcatta atgaatcggc caacgcgcgg
     2881 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2941 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3001 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3061 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3121 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3181 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccc
//
