LOCUS       sequence-326709-        5422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expressio of a GST fusion of Human ISG15 C78SN13Y.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5422)
  AUTHORS   Narasimhan J, Wang M, Fu Z, Klein JM, Haas AL, Kim JJ
  TITLE     Crystal structure of the interferon-induced ubiquitin-like protein 
            ISG15.
  JOURNAL   J Biol Chem. 2005 Jul 22. 280(29):27356-65.
  PUBMED    15917233
REFERENCE   2  (bases 1 to 5422)
  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: "J Biol 
            Chem. 2005 Jul 22. 280(29):27356-65."
COMMENT     Sequence Label: pGEX4T1-ISG15 C78SN13Y
FEATURES             Location/Qualifiers
     source          1..5422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(934..939,940..957,958..964)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    972..988
                     /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     996..1012
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     996..1012
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1008..1535
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1028..1045)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1028..1044)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1037..1059)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1637..1642,1643..1658,1659..1665)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1673..1689
                     /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             1712..2365
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2323..2345
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2372..2389
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2926..2948)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3093..3111)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3179..3283
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3284..3352,3353..4144)
                     /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     complement(3502..3521)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4315..4903
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4804..4823
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5057..5074
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5147..5224
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5225..5422,1..885)
                     /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(5244..5263)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc
       61 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc
      121 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat
      181 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta
      241 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt
      301 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg
      361 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
      421 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
      481 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
      541 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
      601 gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta
      661 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
      721 ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga
      781 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
      841 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
      901 tgtaagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
      961 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     1021 cggattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     1081 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     1141 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt gcctggtttc
     1201 cggcaccaga agcggtgccg gaaagctggc tggagtgcga tcttcctgag gccgatactg
     1261 tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc gcccatctac accaacgtga
     1321 cctatcccat tacggtcaat ccgccgtttg ttcccacgga gaatccgacg ggttgttact
     1381 cgctcacatt taatgttgat gaaagctggc tacaggaagg ccagacgcga attatttttg
     1441 atggcgttgg aattagctta tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc
     1501 catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca
     1561 aggcgcactc ccgttctgga taatgttttt tgcgccgaca tcataacggt tctggcaaat
     1621 attctgaaat gagctgttga caattaatca tcggctcgta taatgtgtgg aattgtgagc
     1681 ggataacaat ttcacacagg aaacagtatt catgtcccct atactaggtt attggaaaat
     1741 taagggcctt gtgcaaccca ctcgacttct tttggaatat cttgaagaaa aatatgaaga
     1801 gcatttgtat gagcgcgatg aaggtgataa atggcgaaac aaaaagtttg aattgggttt
     1861 ggagtttccc aatcttcctt attatattga tggtgatgtt aaattaacac agtctatggc
     1921 catcatacgt tatatagctg acaagcacaa catgttgggt ggttgtccaa aagagcgtgc
     1981 agagatttca atgcttgaag gagcggtttt ggatattaga tacggtgttt cgagaattgc
     2041 atatagtaaa gactttgaaa ctctcaaagt tgattttctt agcaagctac ctgaaatgct
     2101 gaaaatgttc gaagatcgtt tatgtcataa aacatattta aatggtgatc atgtaaccca
     2161 tcctgacttc atgttgtatg acgctcttga tgttgtttta tacatggacc caatgtgcct
     2221 ggatgcgttc ccaaaattag tttgttttaa aaaacgtatt gaagctatcc cacaaattga
     2281 taagtacttg aaatccagca agtatatagc atggcctttg cagggctggc aagccacgtt
     2341 tggtggtggc gaccatcctc caaaatcgga tctggttccg cgtggatccg gctgggacct
     2401 gacggtgaag atgctggcgg gctacgaatt ccaggtgtcc ctgagcagct ccatgtcggt
     2461 gtcagagctg aaggcgcaga tcacccagaa gatcggcgtg cacgccttcc agcagcgtct
     2521 ggctgtccac ccgagcggtg tggcgctgca ggacagggtc ccccttgcca gccagggcct
     2581 gggccccggc agcacggtcc tgctggtggt cgacaaatcc gacgaacctc tgagcatcct
     2641 ggtgaggaat aacaagggcc gcagcagcac ctacgaggtg cggctgacgc agaccgtggc
     2701 ccacctgaag cagcaagtga gcgggctgga gggtgtgcag gacgacctgt tctggctgac
     2761 cttcgagggg aagcccctgg aggaccagct cccgctgggg gagtacggcc tcaagcccct
     2821 gagcaccgtg ttcatgaatc tgcgcctgcg gggaggctaa ctcgagcggc cgcatcgtga
     2881 ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     2941 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     3001 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     3061 tagcggagtg tataattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     3121 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3181 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3241 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3301 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3361 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3421 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3481 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     3541 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3601 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3661 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3721 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3781 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     3841 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3901 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3961 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4021 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4081 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4141 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4201 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4261 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4321 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4381 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4441 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     4501 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4561 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4621 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4681 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4741 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4801 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4861 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4921 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4981 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     5041 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     5101 ttttctcctt acgcatctgt gcggtatttc acaccgcata aattccgaca ccatcgaatg
     5161 gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt
     5221 gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac
     5281 cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga
     5341 agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa
     5401 acagtcgttg ctgattggcg tt
//
