LOCUS       sequence-422815-        4519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of the kinase domain of cAbl with a 
            TEV-cleavable N-terminal 6xHis affinity tag; For in vitro tyrosine 
            phosphorylation in peptides/proteins and for kinase specificity 
            screenings.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4519)
  AUTHORS   Shah NH, Lobel M, Weiss A, Kuriyan J
  TITLE     Fine-tuning of substrate preferences of the Src-family kinase Lck 
            revealed through a high-throughput specificity screen.
  JOURNAL   Elife. 2018 Mar 16;7. pii: 35190. doi: 10.7554/eLife.35190.
  PUBMED    29547119
REFERENCE   2  (bases 1 to 4519)
  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"; journalName: "Elife."; 
            date: "2018-03-16"; volume: "7. pii: 35190. doi"; pages: " 
            10.7554/eLife.35190"
COMMENT     Sequence Label: pET23a-H6-TEV-cAbl
FEATURES             Location/Qualifiers
     source          1..4519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     448..467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     701..718
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    733..872
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(798..817)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     958..980
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(974..1165)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1674..1693
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1674..1692
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1724..1746
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1765..1782
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1804..1824
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2693..2710
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2763..2781)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2777..2824
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2861..3316
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2948..2967)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3158..3179
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3342..3446
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3447..3515,3516..4307)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3665..3684)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4478..4519,1..547)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
       61 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
      121 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      181 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      241 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      301 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      361 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      421 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      481 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      541 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      601 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
      661 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      721 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
      781 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
      841 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
      901 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
      961 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     1021 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     1081 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     1141 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     1201 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     1261 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     1321 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
     1381 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
     1441 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
     1501 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
     1561 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     1621 tgcgcacccg tggccaggac ccaacgctgc ccgagatctc gatcccgcga aattaatacg
     1681 actcactata gggagaccac aacggtttcc ctctagaaat aattttgttt aactttaaga
     1741 aggagatata ccatgggcag cagccatcat catcatcatc acgattacga tatcccaacg
     1801 accgaaaacc tttacttcca gggccatatg tacgacaagt gggaaatgga gcgcaccgac
     1861 atcaccatga agcacaagtt gggtggaggc cagtacgggg aggtgtacga gggcgtttgg
     1921 aagaagtaca gcctcactgt ggccgtgaag accttgaagg aggacaccat ggaggtggag
     1981 gagttcctga aggaagcggc ggtgatgaag gagatcaaac accctaacct ggtgcagctg
     2041 ctaggggtgt gtacccggga accaccattc tacataatca ctgagttcat gacctatggg
     2101 aacctgctgg actacctgag ggagtgtaac cggcaggagg tgagcgccgt ggtactgctc
     2161 tacatggcca cacagatctc atcagccatg gagtacttgg agaagaagaa cttcatccac
     2221 agagaccttg ctgcccggaa ctgcctggta ggggaaaacc acttggtgaa ggtggctgat
     2281 tttggcctga gcaggttgat gacaggggac acctacacgg cccatgctgg agccaaattc
     2341 cccatcaaat ggaccgcacc tgagagcctg gcctacaaca agttctccat caagtcggac
     2401 gtgtgggcat ttggagtatt gctctgggag attgctacct atggcatgtc accttacccg
     2461 ggaattgacc tgtctcaggt ttatgagctg ctggaaaaag actaccgcat ggagcgccct
     2521 gaaggctgcc cggagaaggt ctacgagctc atgcgagcat gttggcagtg gaacccctct
     2581 gaccggccct cctttgctga aatccaccaa gcctttgaaa ccatgttcca ggaatccagt
     2641 atctagggat ccgaattcga gctccgtcga caagcttgcg gccgcactcg agcaccacca
     2701 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     2761 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     2821 tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca
     2881 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2941 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3001 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3061 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3121 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3181 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3241 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     3301 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3361 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3421 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3481 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3541 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3601 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3661 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3721 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3781 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3841 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3901 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3961 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     4021 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4081 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4141 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4201 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4261 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4321 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4381 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4441 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4501 taatctgctg cttgcaaac
//
