LOCUS       sequence-305763-        5971 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 5971)
  AUTHORS   Tiwari S, Weissman AM
  TITLE     Endoplasmic reticulum (ER)-associated degradation of T cell receptor
            subunits. Involvement of ER-associated ubiquitin-conjugating enzymes
            (E2s).
  JOURNAL   J Biol Chem. 2001 May 11. 276(19):16193-200.
  PUBMED    11278356
REFERENCE   2  (bases 1 to 5971)
  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. 2001 May 11. 276(19):16193-200."
COMMENT     Sequence Label: pGEX-KG-MmUbc7/2783
FEATURES             Location/Qualifiers
     source          1..5971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..24,25..31)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    39..55
                     /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     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     63..79
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             75..602
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(95..112)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(95..111)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(104..126)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(704..709,710..725,726..732)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    740..756
                     /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             779..1432
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1390..1412
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1439..1456
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2542..2564)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2709..2727)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2795..2899
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2900..2968,2969..3760)
                     /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(3118..3137)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3931..4519
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4420..4439
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4673..4690
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4763..4840
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4841..5923
                     /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(4860..4879)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
       61 cacaggaaac agctatgacc atgattacgg attcactggc cgtcgtttta caacgtcgtg
      121 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
      181 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
      241 atggcgaatg gcgctttgcc tggtttccgg caccagaagc ggtgccggaa agctggctgg
      301 agtgcgatct tcctgaggcc gatactgtcg tcgtcccctc aaactggcag atgcacggtt
      361 acgatgcgcc catctacacc aacgtgacct atcccattac ggtcaatccg ccgtttgttc
      421 ccacggagaa tccgacgggt tgttactcgc tcacatttaa tgttgatgaa agctggctac
      481 aggaaggcca gacgcgaatt atttttgatg gcgttggaat tagcttatcg actgcacggt
      541 gcaccaatgc ttctggcgtc aggcagccat cggaagctgt ggtatggctg tgcaggtcgt
      601 aaatcactgc ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgttttttgc
      661 gccgacatca taacggttct ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcg
      721 gctcgtataa tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagtattcat
      781 gtcccctata ctaggttatt ggaaaattaa gggccttgtg caacccactc gacttctttt
      841 ggaatatctt gaagaaaaat atgaagagca tttgtatgag cgcgatgaag gtgataaatg
      901 gcgaaacaaa aagtttgaat tgggtttgga gtttcccaat cttccttatt atattgatgg
      961 tgatgttaaa ttaacacagt ctatggccat catacgttat atagctgaca agcacaacat
     1021 gttgggtggt tgtccaaaag agcgtgcaga gatttcaatg cttgaaggag cggttttgga
     1081 tattagatac ggtgtttcga gaattgcata tagtaaagac tttgaaactc tcaaagttga
     1141 ttttcttagc aagctacctg aaatgctgaa aatgttcgaa gatcgtttat gtcataaaac
     1201 atatttaaat ggtgatcatg taacccatcc tgacttcatg ttgtatgacg ctcttgatgt
     1261 tgttttatac atggacccaa tgtgcctgga tgcgttccca aaattagttt gttttaaaaa
     1321 acgtattgaa gctatcccac aaattgataa gtacttgaaa tccagcaagt atatagcatg
     1381 gcctttgcag ggctggcaag ccacgtttgg tggtggcgac catcctccaa aatcggatct
     1441 ggttccgcgt ggatccccgg gaatttccgg tggtggtggt ggaattcggc acgaggaacg
     1501 caggttccgc ggtatggcgg ggacggcgtt gaagaggctg atggccgagt ataagcaatt
     1561 aaccctgaat cctccagaag gaatcgtggc aggccccatg aatgaagaga atttttttga
     1621 atgggaggca ttgatcatgg gccctgaaga cacttgtttt gagtttggtg tttttcccgc
     1681 catcctgagt ttcccacttg actacccctt gagccctccg aagatgagat tcacctgtga
     1741 gatgttccat cccaacatct atcctgatgg cagagtctgc atctccatcc tgcatgctcc
     1801 aggtgatgat cccatgggtt atgagagcag tgccgagcgg tggagcccag tgcagagtgt
     1861 ggagaagatc ctgctttctg ttgtgagcat gctggcagag cccaacgatg agagtggagc
     1921 aaacgtagat gcttccaaga tgtggcggga tgaccgggag cagttctaca agatcgccaa
     1981 gcagattgtc cagaagtctc tggggctctg aggctacccc tgtgcacact gagcactctg
     2041 ctcgagaacc tgtggcgaca gcgctgccat gccaacacca acacaggcag tgcagagcca
     2101 cagcccccgt tgtttcttcc tttccaccca agccaggcag ctccctagat catgactggg
     2161 ataatgacag ctgctgcgct gctaaagact cctcctgctg tgttgtgtgc gcacctcagg
     2221 gagggcccag gctgtcctca ctacctctct cagccatggg gtcctcacct ctgctccttc
     2281 ctgcagtgcc gagggcctgt cctgttgtct gccagggact ggatactaat gctgctccct
     2341 cttgggacat ggggccttgg aaaagcagtg caggcctgtt gtccctgcct ctcagttccc
     2401 aggtgtccca ggaagcattt tgagcttctt ttgacacttt gcacttagaa gcctaggttg
     2461 cctttggcat ggtgacagaa gcttaattca tcgtgactga ctgacgatct gcctcgcgcg
     2521 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     2581 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     2641 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata attcttgaag
     2701 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     2761 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2821 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2881 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2941 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3001 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3061 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3121 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     3181 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3241 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3301 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3361 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3421 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3481 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3541 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3601 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3661 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3721 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3781 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3841 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3901 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3961 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4021 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     4081 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4141 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4201 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4261 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4321 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4381 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4441 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4501 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4561 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4621 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4681 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4741 tatttcacac cgcataaatt ccgacaccat cgaatggcgc aaaacctttc gcggtatggc
     4801 atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata
     4861 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
     4921 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
     4981 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
     5041 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
     5101 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
     5161 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
     5221 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
     5281 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac
     5341 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
     5401 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
     5461 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
     5521 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
     5581 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
     5641 tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac
     5701 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
     5761 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
     5821 aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     5881 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     5941 taagttagct cactcattag gcaccccagg c
//
