LOCUS       sequence-211936-        5485 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 5485)
  AUTHORS   Bruderer RM, Brasseur C, Meyer HH
  TITLE     The AAA ATPase p97/VCP interacts with its alternative co-factors, 
            Ufd1-Npl4 and p47, through a common bipartite binding mechanism.
  JOURNAL   J Biol Chem. 2004 Nov 26. 279(48):49609-16.
  PUBMED    15371428
REFERENCE   2  (bases 1 to 5485)
  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. 2004 Nov 26. 279(48):49609-16."
COMMENT     Sequence Label: pTrcHis-p47
FEATURES             Location/Qualifiers
     source          1..5485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(193..198,199..215,216..222)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    230..246
                     /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     235..257
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    263..332
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     CDS             383..409
                     /codon_start=1
                     /product="synthetic cistron containing a ribosome binding 
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /label=minicistron
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
                     /translation="MYRLNKEE"
     CDS             425..442
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             446..478
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(482..490,491..505)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(1669..1686)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1839..1925
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2017..2044
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2063..2154
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2155..2223,2224..3015)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2373..2392)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3186..3774
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3675..3694
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3928..3945
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3960..4100
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4026..4045)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4186..4208
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4286..4363
                     /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             4364..5446
                     /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(4383..4402)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagcgccgct gagaaaaagc gaagcggcac tgctctttaa
      301 caatttatca gacaatctgt gtgggcactc gaccggaatt atcgattaac tttattatta
      361 aaaattaaag aggtatatat taatgtatcg attaaataag gaggaataaa ccatgggggg
      421 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg
      481 ggatctgtac gacgatgacg ataaggatcc ggcggaggag cggcaggacg cgttgaggga
      541 gttcgtggcg gtgacgggcg ctgaggagga ccgggcccgt ttcttcttgg agtcagctgg
      601 ctgggacctg cagattgcac ttgcaagctt ttatgaggat ggaggggatg aagacattgt
      661 gaccatttca caggcaaccc caagttcagt gtccagaggc acagctccta gtgataatag
      721 agtaacatcc ttcagagacc tcattcatga ccaagatgaa gaggaagagg aagaagaggg
      781 acagaggttt tatgctgggg gctcagagag aagtggacag cagattgttg gtcctccccg
      841 gaagaaaagt cccaacgagc tggtggatga tctgtttaag ggtgccaaag agcatggggc
      901 tgtagctgtg gaacgagtga ccaagagccc tggagagacc agtaaaccga gaccatttgc
      961 aggagggggc taccgccttg gagcagcacc agaagaggag tctgcctatg tggcaggaga
     1021 aaggaggcgg cattcaggcc aggatgttca tgtagtgcta aagctctgga aaactggatt
     1081 cagcctagac aatggtgacc ttagaagcta ccaagaccca tccaatgccc agtttctgga
     1141 gtctatccgt agaggggagg ttcctgcaga gcttcggagg ttagctcacg gtgggcaggt
     1201 gaacttggat atggaggatc atcgggacga ggacttcgtg aagcccaagg gagctttcaa
     1261 agccttcact ggcgagggcc agaaactagg cagcacggcc ccgcaggttt taaacaccag
     1321 ctctccagcc cagcaagcag aaaatgaagc caaagctagc tcgtccatct taatcaatga
     1381 ggcagaacct accacgaaca tccaaatccg actggcagat ggcgggagac tggtgcagaa
     1441 attcaaccac agccacagga tcagcgacat ccggctcttc atcgtggatg cacggccggc
     1501 tatggctgcc accagttttg tccttatgac taccttccct aacaaagagc tggctgatga
     1561 gaaccaaacc ctgaaggaag ccaatcttct caacgctgtc atcgtgcagc ggttaacata
     1621 accgaattcg aagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat
     1681 taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt
     1741 ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt
     1801 ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
     1861 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     1921 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
     1981 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
     2041 tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg
     2101 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     2161 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2221 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2281 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     2341 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     2401 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2461 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2521 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     2581 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2641 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2701 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2761 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2821 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2881 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2941 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3001 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     3061 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3121 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3181 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3241 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3301 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     3361 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3421 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3481 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3541 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3601 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3661 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3721 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3781 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3841 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3901 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3961 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     4021 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     4081 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     4141 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     4201 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gcagatcaat tcgcgcgcga
     4261 aggcgaagcg gcatgcattt acgttgacac catcgaatgg tgcaaaacct ttcgcggtat
     4321 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
     4381 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
     4441 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
     4501 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
     4561 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
     4621 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
     4681 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
     4741 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
     4801 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
     4861 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
     4921 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
     4981 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
     5041 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
     5101 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
     5161 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
     5221 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
     5281 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
     5341 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5401 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5461 atgtaagtta gcgcgaattg atctg
//
