LOCUS       sequence-317088-        5287 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 5287)
  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 5287)
  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 (773-908)
FEATURES             Location/Qualifiers
     source          1..5287
                     /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(2209..2226)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2379..2465
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2557..2584
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2603..2694
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2695..2763,2764..3555)
                     /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(2913..2932)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3726..4314
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4215..4234
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4468..4485
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4562..4990)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4672..4693)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4884..4903
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(5028..5047)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5220..5242
                     /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 ccgggatgaa caagctatct agaaaagcta ttcataagat
     1621 cgtggatatt cgtttgaagg aaattgaaga gagattcgag caaaatgata aacattacaa
     1681 gttgaattta actcaagagg ccaaggactt cttggccaaa tatggttatt ccgatgatat
     1741 gggtgcacgt ccactgaaca ggttaattca aaacgaaatt ttgaacaaac tggcactaag
     1801 gatcttaaag aatgaaatca aggataagga aactgtcaat gtcgtcttga agaagggtaa
     1861 atctcgtgat gaaaatgttc ctgaggaagc tgaagaatgt ctggaagttc taccaaatca
     1921 cgaagctact ataggggctg acacgttagg tgatgacgat aatgaggaca gtatggaaat
     1981 tgatgatgac ctagattaat ttaatatagt gtgattttta aaaactttcg aacaagaatc
     2041 agtaatataa tatatataat taataaaact aatggaattt gtttaattga acttgacacc
     2101 cgagcaaaag agctcactag tcgcggccgc tttcgaatct agagcctgca gtctcgaggc
     2161 atgcggtacc aagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat
     2221 taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt
     2281 ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt
     2341 ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
     2401 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     2461 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
     2521 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
     2581 tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg
     2641 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     2701 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2761 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2821 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     2881 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     2941 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3001 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3061 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     3121 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3181 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3241 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3301 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3361 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     3421 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3481 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3541 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     3601 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3661 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3721 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3781 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3841 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     3901 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3961 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4021 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4081 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4141 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4201 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4261 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4321 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     4381 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     4441 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     4501 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat aattttgtta
     4561 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4621 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4681 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4741 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4801 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4861 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4921 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4981 cagggcgcgt cccattcgcc attcaggctg ctatggtgca ctctcagtac aatctgctct
     5041 gatgccgcat agttaagcca gtaccagtca cgtagcgata tcgctacgtg actggtacac
     5101 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     5161 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5221 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagcagat
     5281 caattcg
//
