LOCUS       sequence-65597-e        5763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5763)
  AUTHORS   Machida K, Thompson CM, Dierck K, Jablonowski K, Karkkainen S, Liu 
            B, Zhang H, Nash PD, Newman DK, Nollau P, Pawson T, Renkema GH, 
            Saksela K, Schiller MR, Shin DG, Mayer BJ
  TITLE     High-throughput phosphotyrosine profiling using SH2 domains.
  JOURNAL   Mol Cell. 2007 Jun 22;26(6):899-915.
  PUBMED    17588523
REFERENCE   2  (bases 1 to 5763)
  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: "Mol Cell.";
            date: "2007-06-22"; volume: "26(6)"; pages: "899-915"
COMMENT     Sequence Label: pAC4 Grb14-PIRSH2
FEATURES             Location/Qualifiers
     source          1..5763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(193..198,199..215,216..222)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    230..246
                     /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        join(470..475,476..492,493..499)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    507..523
                     /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             556..1209
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1167..1189
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1216..1233
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1852..1896
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1946..1963)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2116..2202
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2294..2321
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2341..2432
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2433..2501,2502..3293)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2651..2670)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3464..4052
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3953..3972
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4206..4223
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4238..4378
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4304..4323)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4464..4486
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4564..4641
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4642..5724
                     /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(4661..4680)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagaccatgg agctcgagtt tgacagctta tcatcgactg
      301 cacggtgcac caatgcttct ggcgtcaggc agccatcgga agctgtggta tggctgtgca
      361 ggtcgtaaat cactgcataa ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt
      421 ttttgcgccg acatcataac ggttctggca aatattctga aatgagctgt tgacaattaa
      481 tcatccggct cgtataatgt gtggaattgt gagcggataa caatttcaca caggaaacag
      541 accatggagc tcgagatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
      601 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
      661 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
      721 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
      781 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
      841 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
      901 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
      961 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
     1021 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
     1081 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
     1141 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
     1201 cctccaaaat cggatctggt tccgcgtgga tccaagtatg gcatgcagct gtaccagaat
     1261 tatatgcatc catatcaagg tagaagtggc tgcagttcac agagcatatc acctatgaga
     1321 agtatatcag agaattccct ggtagcaatg gacttctcag gccagaaaag cagagttata
     1381 gaaaatccca ctgaagccct ttcagttgcg gttgaagaag gactcgcttg gaggaaaaaa
     1441 ggatgtttac gcctgggcac tcacggtagc cccactgcct cttcacagag ctctgccaca
     1501 aacatggcta tccaccggtc ccagccatgg tttcaccaca aaatttctag agatgaggct
     1561 cagcgattga ttattcagca aggacttgtg gatggagttt tcttggtacg ggatagtcag
     1621 agtaacccca aaactttcgt actgtcaatg agtcatggac aaaaaataaa gcactttcaa
     1681 attataccag tagaagatga cggtgaaatg ttccacacac tggatgatgg ccacacaaga
     1741 tttacagatc taatacagct ggtggagttc tatcaactca ataagggcgt tcttccttgc
     1801 aagttgaaac attattgtgc taggattgct ctcgaaagct tgcttggtgg cggtctgaac
     1861 gacatcttcg aggctcagaa aatcgaatgg cacgaataat taattaagag cttggctgtt
     1921 ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc
     1981 tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
     2041 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag
     2101 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     2161 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     2221 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     2281 catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt
     2341 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2401 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2461 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2521 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2581 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2641 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     2701 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2761 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2821 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2881 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2941 ccataccaaa cgacgagcgt gacaccacga tgcctacagc aatggcaaca acgttgcgca
     3001 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3061 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3121 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3181 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3241 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3301 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3361 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3421 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3481 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3541 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3601 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3661 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3721 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3781 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3841 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3901 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3961 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4021 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4081 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4141 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4201 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     4261 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     4321 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     4381 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4441 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4501 accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc atgcatttac
     4561 gttgacacca tcgaatggtg caaaaccttt cgcggtatgg catgatagcg cccggaagag
     4621 agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc
     4681 ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa
     4741 acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca
     4801 caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg
     4861 cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc
     4921 gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat
     4981 cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc
     5041 attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag
     5101 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat
     5161 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg
     5221 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
     5281 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
     5341 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
     5401 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac
     5461 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
     5521 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
     5581 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat
     5641 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     5701 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc gcgaattgat
     5761 ctg
//
