LOCUS       sequence-215949-        7887 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of SERPINA5_Serpin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7887)
  AUTHORS   Teyra J, Kelil A, Jain S, Helmy M, Jajodia R, Hooda Y, Gu J, D'Cruz 
            AA, Nicholson SE, Min J, Sudol M, Kim PM, Bader GD, Sidhu SS
  TITLE     Large-scale survey and database of high affinity ligands for peptide
            recognition modules.
  JOURNAL   Mol Syst Biol. 2020 Dec;16(12):e9310. doi: 10.15252/msb.20199310.
  PUBMED    33438817
REFERENCE   2  (bases 1 to 7887)
  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 Syst 
            Biol."; date: "2020-12"; volume: "16(12):e9310. doi"; pages: " 
            10.15252/msb.20199310"
COMMENT     Sequence Label: pHH0103_SERPINA5_Serpin
FEATURES             Location/Qualifiers
     source          1..7887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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     42..64
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             138..155
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             171..824
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     782..804
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             831..854
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             867..884
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      2082..2129
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(3722..4177)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3859..3880)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4071..4090
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4219..4241)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4327..4467
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4382..4401
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4482..4499)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4653..5240)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5411..6202,6203..6271))
                     /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     6034..6053
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6272..6376)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6444..6462
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6485..6562
                     /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             6563..7645
                     /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(6582..6601)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag ccagtccgtt taggtgtttt cacgagcact tcaccaacaa ggaccataga
      121 ttatgaaaat cgaagaacac catcaccatc accattccag cggtaagctt atgtccccta
      181 tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt ttggaatatc
      241 ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa tggcgaaaca
      301 aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat ggtgatgtta
      361 aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac atgttgggtg
      421 gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg gatattagat
      481 acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt gattttctta
      541 gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa acatatttaa
      601 atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat gttgttttat
      661 acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa aaacgtattg
      721 aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca tggcctttgc
      781 agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcggat ctggaagttc
      841 tgttccaggg gcccctgtcc agcggtctgg ttccgcgtgg ttccggtacc gcggcccagc
      901 cggcccatcg tcatcacccg cgcgaaatga aaaaacgtgt tgaagatctg cacgttggcg
      961 cgaccgtcgc cccgagctct cgtcgcgatt ttacgttcga cctgtatcgt gcactggcta
     1021 gtgcggcccc gagccagtct attttcttta gccctgtgag tatctccatg tcactggcaa
     1081 tgctgagtct gggcgctggt agttccacca aaatgcagat tctggaaggc ctgggtctga
     1141 acctgcaaaa atcatcggaa aaagaactgc atcgtggttt tcagcaactg ctgcaggaac
     1201 tgaaccaacc gcgcgatggc tttcagctgt ctctgggtaa tgcgctgttc accgacctgg
     1261 tggttgatct gcaagacacc ttcgtttcag cgatgaaaac gctgtatctg gccgatacct
     1321 ttccgacgaa cttccgtgac tcggcaggtg caatgaaaca gattaatgat tacgttgcca
     1381 aacaaaccaa aggtaaaatc gtcgatctgc tgaaaaacct ggacagcaat gcagtcgtga
     1441 tcatggtgaa ctacatcttt ttcaaagcta aatgggaaac ctcttttaat cataaaggca
     1501 cgcaggaaca agatttctac gtcacctcag aaacggttgt ccgtgtgccg atgatgtcgc
     1561 gcgaagatca gtatcactac ctgctggacc gtaacctgag ctgccgtgtg gttggtgtcc
     1621 cgtaccaggg taatgcaacc gctctgttta ttctgccgag cgaaggcaaa atgcagcaag
     1681 tggaaaacgg tctgtctgaa aaaacgctgc gtaaatggct gaaaatgttt aaaaaacgcc
     1741 agctggaact gtatctgccg aaattcagta ttgaaggctc ctaccaactg gaaaaagtgc
     1801 tgccgtcact gggtatctcg aatgttttta ccagccatgc agatctgtcg ggcattagca
     1861 accactctaa tatccaggtg tctgaaatgg ttcacaaagc cgtcgtggaa gttgacgaat
     1921 ccggtacccg tgcagctgcg gccaccggta cgatctttac gttccgtagc gcgcgcctga
     1981 actctcagcg tctggtcttt aatcgcccgt ttctgatgtt cattgtggat aacaatatcc
     2041 tgttcctggg taaagttaat cgcccggcgg ccgcataata aaccgataca attaaaggct
     2101 ccttttggag cctttttttt ggagattttc aacgtgaaaa aattattatt cgcaattcct
     2161 ttagttgttc ctttctattc tcactccgct gaaactgttg aaagttgttt agcaaaaccc
     2221 catacagaaa attcatttac taacgtctgg aaagacgaca aaactttaga tcgttacgct
     2281 aactatgagg gctgtctgtg gaatgctaca ggcgttgtag tttgtactgg tgacgaaact
     2341 cagtgtctag ctagagtggc ggtggctctg gttccggtga ttttgattat gaaaagatgg
     2401 caaacgctaa taagggggct atgaccgaaa atgccgatga aaacgcgcta cagtctgacg
     2461 ctaaaggcaa acttgattct gtcgctactg attacggtgc tgctatcgat ggtttcattg
     2521 gtgacgtttc cggccttgct aatggtaatg gtgctactgg tgattttgct ggctctaatt
     2581 cccaaatggc tcaagtcggt gacggtgata attcaccttt aatgaataat ttccgtcaat
     2641 atttaccttc cctccctcaa tcggttgaat gtcgcccttt tgtctttggc gctggtaaac
     2701 catatgaatt ttctattgat tgtgacaaaa taaacttatt ccgtggtgtc tttgcgtttc
     2761 ttttatatgt tgccaccttt atgtatgtat tttctacgtt tgctaacata ctgcgtaata
     2821 aggagtctta atcatgccag ttcttttggc tagcgccgcc ctataccttg tctgcctccc
     2881 cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc tgaatggaag ccggcggcac
     2941 ctcgctaacg gattcaccac tccaagaatt ggagccaatc aattcttgcg gagaactgtg
     3001 aatgcgcaaa ccaacccttg gcagaacata tccatcgcgt ccgccatctc cagcagccgc
     3061 acgcggcgca tctcgggcag cgttgggtcc tggccacggg tgcgcatgat cgtgctcctg
     3121 tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg
     3181 atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca
     3241 tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc
     3301 accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca
     3361 tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc
     3421 cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta
     3481 acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat
     3541 cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc
     3601 tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa
     3661 caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg caggatccgc
     3721 gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3781 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3841 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3901 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3961 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     4021 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     4081 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     4141 acacccgccg cgcttaatgc gccgctacag ggcgcgtcgc ggatcctgcc tcgcgcgttt
     4201 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     4261 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     4321 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
     4381 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     4441 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     4501 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4561 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4621 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4681 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4741 cagggtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4801 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4861 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4921 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4981 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5041 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     5101 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5161 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5221 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5281 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5341 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5401 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5461 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5521 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5581 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5641 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5701 gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta
     5761 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5821 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5881 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5941 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6001 gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt
     6061 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6121 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6181 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6241 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6301 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6361 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     6421 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaagaat
     6481 tcccgacacc atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga
     6541 gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc
     6601 cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa
     6661 aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc
     6721 acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct
     6781 gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag
     6841 cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa
     6901 tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc
     6961 cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca
     7021 gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca
     7081 tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc
     7141 ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat
     7201 agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct
     7261 gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc
     7321 aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata
     7381 cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca aacaggattt
     7441 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt
     7501 gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa
     7561 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     7621 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt
     7681 aggcacaatt ctcatgtttg acagcttatc atcgactgca cggtgcacca atgcttctgg
     7741 cgtcaggcag ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca ctgcataatt
     7801 cgtgtcgctc aaggcgcact cccgttctgg ataatgtttt ttgcgccgac atcataacgg
     7861 ttctggcaaa tattctgaaa tgagctg
//
