LOCUS       sequence-317080-        5331 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 5331)
  AUTHORS   Cashikar AG, Schirmer EC, Hattendorf DA, Glover JR, Ramakrishnan MS,
            Ware DM, Lindquist SL
  TITLE     Defining a pathway of communication from the C-terminal peptide 
            binding domain to the N-terminal ATPase domain in a AAA protein.
  JOURNAL   Mol Cell 2002 Apr;9(4):751-60.
  PUBMED    11983167
REFERENCE   2  (bases 1 to 5331)
  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: "Mol Cell"; 
            date: "2002-04"; volume: "9(4)"; pages: "751-60"
COMMENT     Sequence Label: pPROEX Hsp104 (361-548)
FEATURES             Location/Qualifiers
     source          1..5331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        33..110
                     /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             111..1193
                     /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(130..149)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    1462..1478
                     /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     1467..1489
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1511..1528
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1550..1570
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2255..2272)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2425..2511
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2603..2630
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2649..2740
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2741..2809,2810..3601)
                     /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(2959..2978)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3772..4360
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4261..4280
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4514..4531
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4608..5035)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4718..4739)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4929..4948
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(5072..5091)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5264..5286
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cgcgcgaagg cgaagcggca tgcatttacg ttgacaccat cgaatggtgc aaaacctttc
       61 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
      121 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
      181 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
      241 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
      301 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
      361 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
      421 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
      481 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
      541 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
      601 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
      661 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
      721 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
      781 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
      841 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
      901 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
      961 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     1021 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
     1081 tgaaaagaaa aaccaccctg gcacccaata cgcaaaccgc ctctccccgc gcgttggccg
     1141 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     1201 gcaattaatg taagttagcg cgaattgatc tggtttgaca gcttatcatc gactgcacgg
     1261 tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg
     1321 taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg
     1381 cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc
     1441 cggtccgtat aatctgtgga attgtgagcg gataacaatt tcacacagga aacagaccat
     1501 gtcgtactac catcaccatc accatcacga ttacgatatc ccaacgaccg aaaacctgta
     1561 ttttcagggc gccatgggat ccgggatgat acatcatggt gtaaggattc tggatagcgc
     1621 cttagtcacc gcggctcaat tagccaagcg ttacttgcca tatagaagat tgccagattc
     1681 tgctttggat ttagttgata tttcttgtgc tggtgtcgcc gtcgcaagag attctaagcc
     1741 agaagaattg gattccaagg aacgtcaatt gcaattgatt caagtagaga taaaagctct
     1801 agagagagat gaagatgccg actccaccac taaagataga ttaaagttag ctaggcagaa
     1861 ggaagcttca ttgcaagaag aattggaacc tctaagacaa cgttacaatg aagaaaagca
     1921 tggccatgaa gaattgacac aagctaaaaa gaaattggat gaactggaaa acaaggccct
     1981 tgatgctgaa cgtagatatg atactgctac cgccgctgat ttaaggtact tcgccatccc
     2041 agatatcaaa aagcaaatcg aaaagcttga agatcaggtt gctgaggaag agagacgtgc
     2101 tggtgccaac tccatgatcc aaaatgtggt cgactcagac accatttctg aataggagct
     2161 cactagtcgc ggccgctttc gaatctagag cctgcagtct cgaggcatgc ggtaccaagc
     2221 ttggctgttt tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca
     2281 gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc
     2341 ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg
     2401 cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     2461 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga
     2521 gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa
     2581 actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta
     2641 caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2701 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     2761 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     2821 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     2881 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     2941 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     3001 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3061 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3121 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3181 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3241 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3301 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3361 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3421 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3481 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3541 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     3601 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     3661 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3721 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3781 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3841 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3901 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3961 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4021 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4081 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4141 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4201 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4261 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4321 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4381 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4441 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     4501 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     4561 tctccttacg catctgtgcg gtatttcaca ccgcataatt ttgttaaaat tcgcgttaaa
     4621 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     4681 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     4741 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     4801 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     4861 tcgaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4921 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     4981 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     5041 tcgccattag gctgctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5101 gccagtacca gtcacgtagc gatatcgcta cgtgactggt acactccgct atcgctacgt
     5161 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     5221 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     5281 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc agatcaattc g
//
