LOCUS       sequence-365322-        5472 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of TSG101 UEV domain (aa 2-145).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5472)
  AUTHORS   Im YJ, Kuo L, Ren X, Burgos PV, Zhao XZ, Liu F, Burke TR, Bonifacino
            JS, Freed EO, Hurley JH
  TITLE     Crystallographic and functional analysis of the ESCRT-I /HIV-1 Gag 
            PTAP interaction.
  JOURNAL   Structure. 2010 Nov 10. 18(11):1536-47.
  PUBMED    21070952
REFERENCE   2  (bases 1 to 5472)
  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: "Structure. 
            2010 Nov 10. 18(11):1536-47."
COMMENT     Sequence Label: pGST2-TSG101 (2-145)
FEATURES             Location/Qualifiers
     source          1..5472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(531..536,537..554,555..561)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    569..585
                     /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     593..609
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     593..609
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             605..1132
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(625..642)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(625..641)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(634..656)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1234..1239,1240..1255,1256..1262)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1270..1286
                     /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             1309..1962
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1920..1942
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1975..1995
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2573..2595)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2740..2758)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2826..2930
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2931..2999,3000..3791)
                     /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(3149..3168)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3962..4550
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4451..4470
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4704..4721
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4794..4871
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(4872..5472,1..482)
                     /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(4891..4910)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
       61 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
      121 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
      181 ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat
      241 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
      301 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg
      361 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
      421 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
      481 gagcgcaacg caattaatgt aagttagctc actcattagg caccccaggc tttacacttt
      541 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
      601 agctatgacc atgattacgg attcactggc cgtcgtttta caacgtcgtg actgggaaaa
      661 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
      721 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
      781 gcgctttgcc tggtttccgg caccagaagc ggtgccggaa agctggctgg agtgcgatct
      841 tcctgaggcc gatactgtcg tcgtcccctc aaactggcag atgcacggtt acgatgcgcc
      901 catctacacc aacgtgacct atcccattac ggtcaatccg ccgtttgttc ccacggagaa
      961 tccgacgggt tgttactcgc tcacatttaa tgttgatgaa agctggctac aggaaggcca
     1021 gacgcgaatt atttttgatg gcgttggaat tagcttatcg actgcacggt gcaccaatgc
     1081 ttctggcgtc aggcagccat cggaagctgt ggtatggctg tgcaggtcgt aaatcactgc
     1141 ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgttttttgc gccgacatca
     1201 taacggttct ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcg gctcgtataa
     1261 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagtattcat gtcccctata
     1321 ctaggttatt ggaaaattaa gggccttgtg caacccactc gacttctttt ggaatatctt
     1381 gaagaaaaat atgaagagca tttgtatgag cgcgatgaag gtgataaatg gcgaaacaaa
     1441 aagtttgaat tgggtttgga gtttcccaat cttccttatt atattgatgg tgatgttaaa
     1501 ttaacacagt ctatggccat catacgttat atagctgaca agcacaacat gttgggtggt
     1561 tgtccaaaag agcgtgcaga gatttcaatg cttgaaggag cggttttgga tattagatac
     1621 ggtgtttcga gaattgcata tagtaaagac tttgaaactc tcaaagttga ttttcttagc
     1681 aagctacctg aaatgctgaa aatgttcgaa gatcgtttat gtcataaaac atatttaaat
     1741 ggtgatcatg taacccatcc tgacttcatg ttgtatgacg ctcttgatgt tgttttatac
     1801 atggacccaa tgtgcctgga tgcgttccca aaattagttt gttttaaaaa acgtattgaa
     1861 gctatcccac aaattgataa gtacttgaaa tccagcaagt atatagcatg gcctttgcag
     1921 ggctggcaag ccacgtttgg tggtggcgac catcctccaa aaatcgatac gaccgaaaac
     1981 ctgtattttc agggcgccat gggatccgcg gtgtcggaga gccagctcaa gaaaatggtg
     2041 tccaagtaca aatacagaga cctaactgta cgtgaaactg tcaatgttat tactctatac
     2101 aaagatctca aacctgtttt ggattcatat gtttttaacg atggcagttc cagggaacta
     2161 atgaacctca ctggaacaat ccctgtgcct tatagaggta atacatacaa tattccaata
     2221 tgcctatggc tactggacac atacccatat aatcccccta tctgttttgt taagcctact
     2281 agttcaatga ctattaaaac aggaaagcat gttgatgcaa atgggaagat atatcttcct
     2341 tatctacatg aatggaaaca cccacagtca gacttgttgg ggcttattca ggtcatgatt
     2401 gtggtatttg gagatgaacc tccagtcttc tctcgtcctt gagaattcaa aggcctacgt
     2461 cgacgagctc actagtcgcg gccgctttcg aatctagagc ctgcagtctc gagcggccgc
     2521 atcgtgactg actgacgatc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
     2581 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     2641 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac
     2701 gtagcgatag cggagtgtat aattcttgaa gacgaaaggg cctcgtgata cgcctatttt
     2761 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     2821 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2881 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2941 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3001 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3061 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3121 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     3181 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3241 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3301 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3361 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3421 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     3481 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3541 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3601 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3661 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3721 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3781 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3841 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3901 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3961 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4021 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4081 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     4141 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4201 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4261 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4321 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4381 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4441 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4501 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4561 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4621 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4681 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     4741 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataaat tccgacacca
     4801 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
     4861 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
     4921 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
     4981 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
     5041 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
     5101 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
     5161 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
     5221 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
     5281 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
     5341 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
     5401 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
     5461 gtctggctgg ct
//
