LOCUS       sequence-140198-        9034 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 9034)
  AUTHORS   Fang S, Ferrone M, Yang C, Jensen JP, Tiwari S, Weissman AM
  TITLE     The tumor autocrine motility factor receptor, gp78, is a ubiquitin 
            protein ligase implicated in degradation from the endoplasmic 
            reticulum.
  JOURNAL   Proc Natl Acad Sci U S A. 2001 Dec 4. 98(25):14422-7.
  PUBMED    11724934
REFERENCE   2  (bases 1 to 9034)
  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: "Proc Natl 
            Acad Sci U S A. 2001 Dec 4. 98(25):14422-7."
COMMENT     Sequence Label: pGEX-KG-Itch/2793
FEATURES             Location/Qualifiers
     source          1..9034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        63..140
                     /gene="lacI"
                     /label=lacI promoter
     CDS             141..1223
                     /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(160..179)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1272..1277,1278..1295,1296..1302)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1310..1326
                     /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     1334..1350
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1334..1350
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1346..1873
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1366..1383)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1366..1382)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1375..1397)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1975..1980,1981..1996,1997..2003)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2011..2027
                     /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             2050..2703
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2661..2683
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2710..2727
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(6876..6898)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7043..7061)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7129..7233
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7234..7302,7303..8094)
                     /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(7452..7471)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8265..8853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8754..8773
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9007..9024
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataaatt
       61 ccgacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
      121 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
      181 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
      241 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
      301 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
      361 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
      421 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
      481 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
      541 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
      601 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
      661 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
      721 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
      781 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
      841 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
      901 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
      961 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
     1021 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     1081 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     1141 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     1201 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
     1261 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     1321 taacaatttc acacaggaaa cagctatgac catgattacg gattcactgg ccgtcgtttt
     1381 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     1441 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     1501 gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg gcaccagaag cggtgccgga
     1561 aagctggctg gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct caaactggca
     1621 gatgcacggt tacgatgcgc ccatctacac caacgtgacc tatcccatta cggtcaatcc
     1681 gccgtttgtt cccacggaga atccgacggg ttgttactcg ctcacattta atgttgatga
     1741 aagctggcta caggaaggcc agacgcgaat tatttttgat ggcgttggaa ttagcttatc
     1801 gactgcacgg tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct
     1861 gtgcaggtcg taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata
     1921 atgttttttg cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca
     1981 attaatcatc ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     2041 acagtattca tgtcccctat actaggttat tggaaaatta agggccttgt gcaacccact
     2101 cgacttcttt tggaatatct tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa
     2161 ggtgataaat ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa tcttccttat
     2221 tatattgatg gtgatgttaa attaacacag tctatggcca tcatacgtta tatagctgac
     2281 aagcacaaca tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga
     2341 gcggttttgg atattagata cggtgtttcg agaattgcat atagtaaaga ctttgaaact
     2401 ctcaaagttg attttcttag caagctacct gaaatgctga aaatgttcga agatcgttta
     2461 tgtcataaaa catatttaaa tggtgatcat gtaacccatc ctgacttcat gttgtatgac
     2521 gctcttgatg ttgttttata catggaccca atgtgcctgg atgcgttccc aaaattagtt
     2581 tgttttaaaa aacgtattga agctatccca caaattgata agtacttgaa atccagcaag
     2641 tatatagcat ggcctttgca gggctggcaa gccacgtttg gtggtggcga ccatcctcca
     2701 aaatcggatc tggttccgcg tggatccccg ggaatttccg gtggtggtgg tggaattcta
     2761 gactccatgg gtagtctgac catgaaatct caacttcaga tcactgtcat ctcagccaaa
     2821 cttaaagaaa ataaaaagaa ttggtttgga ccaagtcctt atgtagaagt cacagtagat
     2881 ggacagtcaa agaagacaga aaaatgcaat aatacaaaca gtcccaagtg gaagcagccc
     2941 ctcacagtta ttgttacccc tacgagtaaa ttatgttttc gtgtgtggag tcaccagacc
     3001 ctgaagtctg atgttttatt gggaactgct ggattagata tttatgaaac attaaagtca
     3061 aacaatatga aacttgaaga agtagttatg actttgcagc ttgtaggtga caaagagcca
     3121 acagagacga tgggagattt gtcagtttgt cttgatgggc tgcaagtaga agctgaggtt
     3181 gttactaacg gtgaaacgtc atgctccgag agtactacac agaatgatga tggctgcaga
     3241 accagagatg atacaagagt gagcacaaat ggatcagagg accctgaggt tgcagcgtca
     3301 ggggaaaaca agagggccaa tgggaacaat tctccgtccc tttcaaatgg tggttttaag
     3361 ccttctagac ctcctagacc ttcgagacca cctccaccca ccccacgaag accagcttct
     3421 gtcaatggct caccatccac gaattctgac agtgatggat ctagtacagg ctccttgcca
     3481 ccaacaaata caaatgtaaa tacaagtaca tctgaaggag caacatctgg attaataatt
     3541 cctcttacta tatctggagg ctcgggccct aggcctctga atactgtaag ccaagctccc
     3601 ctaccacctg ggtgggagca gagagtagac cagcatgggc gtgtttacta tgtagaccat
     3661 gttgaaaagc gaacaacatg ggatagacca gaacctctac ctcctggctg ggaacggcgt
     3721 gtggacaata tgggacgtat ttattatgtt gatcatttca caagaaccac aacatggcag
     3781 aggccaacct tggaatctgt ccggaactat gaacagtggc agctacagcg tagtcagctt
     3841 cagggagcaa tgcagcagtt taaccagaga ttcatctatg ggaatcaaga tttgtttgct
     3901 acatcacaaa acaaagaatt tgatccgctt ggcccgttgc cccctggatg ggagaagaga
     3961 actgatagca atggcagagt gtattttgtc aaccacaaca ctcggattac tcagtgggaa
     4021 gaccccagaa gccaaggtca gttaaatgaa aagcccttac cagaaggctg ggaaatgaga
     4081 ttcacagtgg atggaattcc atattttgtg gaccacaata gaagagcaac tacttatata
     4141 gatccacgaa cgggaaaatc agccttagac aatgggcccc agatagccta tgtgcgggac
     4201 ttcaaggcaa aagttcagta tttccggttc tggtgccagc aactggccat gccacagcat
     4261 ataaagatca cagtgacaag aaaaacattg tttgaggatt cctttcagca gatcatgagc
     4321 ttcagcccgc aagacctgag aagacgtttg tgggtgattt tcccaggaga agaaggttta
     4381 gattatggag gtgtagcaag agaatggttc tttcttttgt cacatgaagt gttgaaccca
     4441 atgtattgcc tgtttgaata tgcagggaag gataactact gcctgcagat aaaccccgct
     4501 tcttacatca atccagacca cctgaaatac tttcgtttta ttggcagatt tattgccatg
     4561 gctctgtttc atgggaaatt catagatacc ggattttctt taccattcta taaacgtatc
     4621 ctgaataaac cagttggact taaggattta gaatctattg atccagaatt ttataattcg
     4681 ctcatctggg ttaaagaaaa caacatcgaa gaatgtggtt tggaaatgta cttctcagtt
     4741 gataaagaaa ttctaggtga aattaagagt catgatttga aaccgaatgg tggcaatatt
     4801 cttgtgacag aagaaaacaa ggaggaatac atcaggatgg tagctgagtg gaggttgtct
     4861 cgaggtgttg aagaacagac acaagctttc tttgagggct ttaatgaaat tcttcctcag
     4921 caatatttgc aatattttga tgcgaaggaa ttagaggttc ttctgtgtgg gatgcaggag
     4981 attgatttga atgactggca gagacatgcc atctaccgcc actacaccag aacaagcaag
     5041 cagatcatgt ggttttggca gtttgttaaa gaaattgata atgagaagag gatgagactt
     5101 ctgcagtttg ttactggaac ctgccgattg ccagtgggag gatttgctga ccttatgggg
     5161 agtaatggac cacagaagtt ctgcatcgaa aaagttggca aagaaaattg gttacccaga
     5221 agccatactt gttttaaccg cctggacctc ccaccttaca agagctatga gcaactgaag
     5281 gaaaagctgt tatttgccat tgaagaaact gaaggatttg gacaagagta attgtttaga
     5341 ccttaagcca ccagaggcca gggacttgtc tgcagtgtct gtccccttcc ttctctgcct
     5401 gttgcacatc ttattgtaaa cttggactgt gactagagag ttacctgaga gtaagtaaat
     5461 taatgttatc gttgagatct tccagtcatt ctggatttct actcaacgtt tccagaaagc
     5521 agatctacaa actgtagtca gaaacttaac atgagattta acacagcgag gaaatttccc
     5581 tcgtcttatt ccactagatt gttcccttaa caacagtttg ctgtatgtcc gaagtctcac
     5641 ttgggcatag gtctcttagt gggtgttgta aaggattctg cagggacatc tcggaaacgc
     5701 aatacacaag gtcttcctgt aaagcctcgc tcagaggtgc aagcttaact cagtgtctgg
     5761 gcacatgagc agctgctgct tgtgctctcc gctcgtccca gcctggcctg actgttgttt
     5821 ttcattcctg gagcatgaat cagtgcacca ttctgatgct attcctggaa gtgatgcagg
     5881 acttgcattt ttctacaaag tagttttgtc tatttgaatt cagggaaaac ttggtccccg
     5941 ccaacatatg accatacctc tcttcattca gtttgtaatt gttctattgc cttgcaaata
     6001 ctaaacttta caaacagtat cagcactgtg gttccttcac gtttttagaa attaacctag
     6061 aattaaaata agttcagatg aattcatcta agatcattct tgattttttt aaatcctaca
     6121 cataaattga agctttatgg tgttgaagta atccaacacc ttagttacat ttcatttttg
     6181 ctaaactatt caagttgcaa ccatttccta acacaaatgg aggtcagtgg tgcagactct
     6241 gcagtcacaa cagcgtccag cagccacaca gctgcgctgt cttcacactc cactcctgac
     6301 acaggagcag ccgtgtctac cctcttacat ggtcctgatc tacaatattt ttctctcccc
     6361 taaaaattgt agatttccct tttctttcca tgactccttt atcttaggct gctcctttag
     6421 taactgatga atgcactttg accttctctg taggtcattt accttggctt agcattagac
     6481 tgtgaggatg gttgctgcca cagactttat gggcagcaag atgaaacctc agccaggtga
     6541 gtccctattg tagccaaatc ccaaaattag ccttatttct ctaatgggga ccttgaaatg
     6601 tgttggtggc atagcataaa gccaagaaca ccgagtgtga caaacccctg tcccatagga
     6661 aagaaagtgt ttctgtgtcc ctagctgctc gtcctgttct catcctggga cagtttgaac
     6721 tcgtgcagct gtgctcctta acactgccat cctcagcaga gacgtgacag acgggttgcc
     6781 tcatgaaaaa gtgatcagtg aagggagagc tcaagcttaa ttcatcgtga ctgactgacg
     6841 atctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     6901 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     6961 gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg
     7021 tataattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     7081 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     7141 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     7201 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     7261 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     7321 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     7381 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     7441 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     7501 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     7561 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     7621 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     7681 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     7741 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
     7801 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     7861 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     7921 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     7981 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     8041 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     8101 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     8161 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     8221 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     8281 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     8341 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     8401 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     8461 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     8521 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     8581 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     8641 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     8701 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     8761 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     8821 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     8881 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     8941 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     9001 gagcgcagcg agtcagtgag cgaggaagcg gaag
//
