LOCUS       sequence-191137-        6969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of human SH3 domain construct 
            ABI1-1/2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6969)
  AUTHORS   Teyra J, Huang H, Jain S, Guan X, Dong A, Liu Y, Tempel W, Min J, 
            Tong Y, Kim PM, Bader GD, Sidhu SS
  TITLE     Comprehensive Analysis of the Human SH3 Domain Family Reveals a Wide
            Variety of Non-canonical Specificities.
  JOURNAL   Structure. 2017 Oct 3;25(10):1598-1610.e3. doi: 
            10.1016/j.str.2017.07.017. Epub 2017 Sep 7.
  PUBMED    28890361
REFERENCE   2  (bases 1 to 6969)
  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: 
            "10.1016/j.str.2017.07.017"; journalName: "Structure"; date: 
            "2017-10-3- 3"; volume: "25"; issue: "10"; pages: "1598-1610.e3"
COMMENT     Sequence Label: pHH0103_ABI1-1/2
FEATURES             Location/Qualifiers
     source          1..6969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..36
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        59..136
                     /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             137..1219
                     /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(156..175)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1462..1467,1468..1483,1484..1490)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1498..1514
                     /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     1503..1525
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1599..1616
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1632..2285
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2243..2265
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2292..2315
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2328..2345
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2625..2672
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(4265..4720)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4402..4423)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4614..4633
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4762..4784)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4870..5010
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4925..4944
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5025..5042)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5196..5783)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5954..6745,6746..6814))
                     /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     6577..6596
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6815..6919)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattcccga
       61 caccatcgaa tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
      121 attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt
      181 ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg
      241 ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca
      301 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc
      361 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt
      421 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct
      481 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc
      541 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc
      601 catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt
      661 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg
      721 tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga
      781 acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga
      841 gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg
      901 cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga
      961 taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct
     1021 gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg
     1081 caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca
     1141 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     1201 actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac
     1261 aattctcatg tttgacagct tatcatcgac tgcacggtgc accaatgctt ctggcgtcag
     1321 gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc
     1381 gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg
     1441 caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg tgtggaattg
     1501 tgagcggata acaatttcac acaggaaaca gccagtccgt ttaggtgttt tcacgagcac
     1561 ttcaccaaca aggaccatag attatgaaaa tcgaagaaca ccatcaccat caccattcca
     1621 gcggtaagct tatgtcccct atactaggtt attggaaaat taagggcctt gtgcaaccca
     1681 ctcgacttct tttggaatat cttgaagaaa aatatgaaga gcatttgtat gagcgcgatg
     1741 aaggtgataa atggcgaaac aaaaagtttg aattgggttt ggagtttccc aatcttcctt
     1801 attatattga tggtgatgtt aaattaacac agtctatggc catcatacgt tatatagctg
     1861 acaagcacaa catgttgggt ggttgtccaa aagagcgtgc agagatttca atgcttgaag
     1921 gagcggtttt ggatattaga tacggtgttt cgagaattgc atatagtaaa gactttgaaa
     1981 ctctcaaagt tgattttctt agcaagctac ctgaaatgct gaaaatgttc gaagatcgtt
     2041 tatgtcataa aacatattta aatggtgatc atgtaaccca tcctgacttc atgttgtatg
     2101 acgctcttga tgttgtttta tacatggacc caatgtgcct ggatgcgttc ccaaaattag
     2161 tttgttttaa aaaacgtatt gaagctatcc cacaaattga taagtacttg aaatccagca
     2221 agtatatagc atggcctttg cagggctggc aagccacgtt tggtggtggc gaccatcctc
     2281 caaaatcgga tctggaagtt ctgttccagg ggcccctgtc cagcggtctg gttccgcgtg
     2341 gttccggtac cgcggcccag ccggccgaag acgaagaagc agcagtggtt cagtacaacg
     2401 acccgtatgc agatggcgac ccggcgtggg ctccgaaaaa ctacattgaa aaagtggttg
     2461 cgatttatga ttacaccaaa gacaaagatg acgaactgag ctttatggaa ggcgccatca
     2521 tctacgttat taagaaaaac gatgacggct ggtacgaagg tgtctgcaac cgtgtgacgg
     2581 gcctgttccc gggtaattat gtcgaatctg cggccgcata ataaaccgat acaattaaag
     2641 gctccttttg gagccttttt tttggagatt ttcaacgtga aaaaattatt attcgcaatt
     2701 cctttagttg ttcctttcta ttctcactcc gctgaaactg ttgaaagttg tttagcaaaa
     2761 ccccatacag aaaattcatt tactaacgtc tggaaagacg acaaaacttt agatcgttac
     2821 gctaactatg agggctgtct gtggaatgct acaggcgttg tagtttgtac tggtgacgaa
     2881 actcagtgtc tagctagagt ggcggtggct ctggttccgg tgattttgat tatgaaaaga
     2941 tggcaaacgc taataagggg gctatgaccg aaaatgccga tgaaaacgcg ctacagtctg
     3001 acgctaaagg caaacttgat tctgtcgcta ctgattacgg tgctgctatc gatggtttca
     3061 ttggtgacgt ttccggcctt gctaatggta atggtgctac tggtgatttt gctggctcta
     3121 attcccaaat ggctcaagtc ggtgacggtg ataattcacc tttaatgaat aatttccgtc
     3181 aatatttacc ttccctccct caatcggttg aatgtcgccc ttttgtcttt ggcgctggta
     3241 aaccatatga attttctatt gattgtgaca aaataaactt attccgtggt gtctttgcgt
     3301 ttcttttata tgttgccacc tttatgtatg tattttctac gtttgctaac atactgcgta
     3361 ataaggagtc ttaatcatgc cagttctttt ggctagcgcc gccctatacc ttgtctgcct
     3421 ccccgcgttg cgtcgcggtg catggagccg ggccacctcg acctgaatgg aagccggcgg
     3481 cacctcgcta acggattcac cactccaaga attggagcca atcaattctt gcggagaact
     3541 gtgaatgcgc aaaccaaccc ttggcagaac atatccatcg cgtccgccat ctccagcagc
     3601 cgcacgcggc gcatctcggg cagcgttggg tcctggccac gggtgcgcat gatcgtgctc
     3661 ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca
     3721 ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca
     3781 acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc
     3841 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct
     3901 acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc
     3961 atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca
     4021 gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga
     4081 aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc
     4141 cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat
     4201 gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcaggatc
     4261 cgcgaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     4321 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     4381 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     4441 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     4501 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     4561 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     4621 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     4681 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cgcggatcct gcctcgcgcg
     4741 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     4801 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     4861 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac
     4921 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     4981 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg
     5041 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5101 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5161 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5221 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5281 taccagggtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5341 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5401 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5461 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5521 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5581 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     5641 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5701 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5761 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5821 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5881 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5941 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6001 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6061 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6121 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6181 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6241 atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg
     6301 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6361 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6421 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6481 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6541 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     6601 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6661 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6721 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6781 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6841 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6901 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     6961 ttattatca
//
