LOCUS       sequence-36-expo        6487 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6487)
  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 6487)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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 (1-575)
FEATURES             Location/Qualifiers
     source          1..6487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     CDS             279..296
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             318..338
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2181..2198)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2351..2437
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2529..2556
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2576..2667
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2668..2736,2737..3528)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2886..2905)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3699..4287
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4188..4207
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4441..4458
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4535..4963)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4645..4666)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4857..4876
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(5001..5020)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5188..5210
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5288..5365
                     /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             5366..6448
                     /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(5385..5404)
                     /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 gtccgtataa tctgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagaccatgt cgtactacca tcaccatcac catcacgatt
      301 acgatatccc aacgaccgaa aacctgtatt ttcagggcgc catgggatcc gggatgaacg
      361 accaaacgca atttacagaa agggctctaa cgattttgac gttggctcaa aaattggctt
      421 cggatcatca acatccacaa ttacaaccta tacatattct agctgccttc attgaaacgc
      481 cagaagatgg atcagtccct tacctacaga atctaattga gaagggccgt tacgactatg
      541 atcttttcaa gaaagtggtt aatagaaatc tagtaagaat tcctcaacag caacctgcac
      601 ctgcggagat aactccaagt tatgctttgg ggaaagtcct tcaagacgct gctaagattc
      661 aaaaacaaca gaaggactca tttatagcgc aagaccatat attgtttgct ctattcaatg
      721 attcgtctat tcagcaaata tttaaggaag ctcaagtaga tattgaggcc atcaagcaac
      781 aagctcttga acttcgtggt aacactagaa ttgactctcg tggcgctgat acgaacacac
      841 ctttggaata tttatcaaag tacgccattg atatgactga gcaggctcgt caaggtaaac
      901 ttgaccctgt catcggccgt gaagaagaaa taagaagcac tattagagtt ttagcaagaa
      961 gaattaagtc caacccatgt ttaattggtg agccaggtat cggtaagacc gctattattg
     1021 aaggtgttgc tcaaagaatc attgacgatg acgttcccac tatcttacaa ggcgctaaat
     1081 tgttcagtct agatttggcc gcattaaccg caggtgctaa atacaaaggt gatttcgaag
     1141 aaagattcaa aggtgttttg aaggaaatcg aagaatcaaa gactctaatt gtgttattca
     1201 ttgatgaaat tcacatgtta atgggtaatg gtaaggacga cgctgctaac atcttgaagc
     1261 cagctttgtc cagaggccaa ttgaaggtca tcggtgccac caccaataac gaatatagat
     1321 ctattgtgga aaaggatggt gcctttgaaa gaagattcca gaaaattgaa gtcgctgaac
     1381 caagtgtgag acaaacagtg gccatattga gaggtctgca accaaagtat gaaatacatc
     1441 atggtgtaag gattctggat agcgccttag tcaccgcggc tcaattagcc aagcgttact
     1501 tgccatatag aagattgcca gattctgctt tggatttagt tgatatttct tgtgctggtg
     1561 tcgccgtcgc aagagattct aagccagaag aattggattc caaggaacgt caattgcaat
     1621 tgattcaagt agagataaaa gctctagaga gagatgaaga tgccgactcc accactaaag
     1681 atagattaaa gttagctagg cagaaggaag cttcattgca agaagaattg gaacctctaa
     1741 gacaacgtta caatgaagaa aagcatggcc atgaagaatt gacacaagct aaaaagaaat
     1801 tggatgaact ggaaaacaag gcccttgatg ctgaacgtag atatgatact gctaccgccg
     1861 ctgatttaag gtacttcgcc atcccagata tcaaaaagca aatcgaaaag cttgaagatc
     1921 aggttgctga ggaagagaga cgtgctggtg ccaactccat gatccaaaat gtggtcgact
     1981 cagacaccat ttctgaaaca gctgcaagat tgactggtat ccctgttaag aagttgtcag
     2041 aatctgaaaa tgaaaaattg attcatatgg aacgtgacta agagctcact agtcgcggcc
     2101 gctttcgaat ctagagcctg cagtctcgag gcatgcggta ccaagcttgg ctgttttggc
     2161 ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata
     2221 aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca
     2281 gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac
     2341 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     2401 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
     2461 tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca
     2521 aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctctttttgt
     2581 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2641 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     2701 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     2761 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     2821 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     2881 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     2941 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3001 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3061 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3121 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3181 ccaaacgacg agcgtgacac cacgatgcct acagcaatgg caacaacgtt gcgcaaacta
     3241 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     3301 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3361 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     3421 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     3481 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     3541 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     3601 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     3661 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3721 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3781 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3841 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3901 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3961 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4021 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4081 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4141 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4201 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4261 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4321 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4381 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     4441 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     4501 ctgtgcggta tttcacaccg cataattttg ttaaaattcg cgttaaattt ttgttaaatc
     4561 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     4621 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     4681 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     4741 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     4801 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     4861 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     4921 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg
     4981 ctgctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtaccag
     5041 tcacgtagcg atatcggagt gtatacactc cgctatcgct acgtgactgg gtcatggctg
     5101 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     5161 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     5221 catcaccgaa acgcgcgagg cagcagatca attcgcgcgc gaaggcgaag cggcatgcat
     5281 ttacgttgac accatcgaat ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga
     5341 agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta
     5401 tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc
     5461 gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt
     5521 ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc
     5581 cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc
     5641 cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca
     5701 caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga
     5761 tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga
     5821 ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga
     5881 gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt
     5941 ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc
     6001 gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat
     6061 gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg
     6121 cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg
     6181 atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga
     6241 ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc
     6301 ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcacc
     6361 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     6421 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagcgcgaat
     6481 tgatctg
//
