LOCUS       sequence-21-expo        5225 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 5225)
  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 5225)
  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: pJC45BS104 Y257W
FEATURES             Location/Qualifiers
     source          1..5225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     65..84
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        65..83
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    84..108
                     /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)."
     RBS             123..145
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             158..187
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             200..211
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3128..3146)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3142..3189
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3233..3251)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3319..3423
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3424..3492,3493..4284)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAISMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3642..3661)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4455..5043
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5197..5214
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg
       61 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tctagaaata
      121 attttgttta actttaagaa ggagatatac catgggccat catcatcatc atcatcatca
      181 tcatcacagc agcggccata tcgaaggtcg tggatccggg atgaacgacc aaacgcaatt
      241 tacagaaagg gctctaacga ttttgacgtt ggctcaaaaa ttggcttcgg atcatcaaca
      301 tccacaatta caacctatac atattctagc tgccttcatt gaaacgccag aagatggatc
      361 agtcccttac ctacagaatc taattgagaa gggccgttac gactatgatc ttttcaagaa
      421 agtggttaat agaaatctag taagaattcc tcaacagcaa cctgcacctg cggagataac
      481 tccaagttat gctttgggga aagtccttca agacgctgct aagattcaaa aacaacagaa
      541 ggactcattt atagcgcaag accatatatt gtttgctcta ttcaatgatt cgtctattca
      601 gcaaatattt aaggaagctc aagtagatat tgaggccatc aagcaacaag ctcttgaact
      661 tcgtggtaac actagaattg actctcgtgg cgctgatacg aacacacctt tggaatattt
      721 atcaaagtac gccattgata tgactgagca ggctcgtcaa ggtaaacttg accctgtcat
      781 cggccgtgaa gaagaaataa gaagcactat tagagtttta gcaagaagaa ttaagtccaa
      841 cccatgttta attggtgagc caggtatcgg taagaccgct attattgaag gtgttgctca
      901 aagaatcatt gacgatgacg ttcccactat cttacaaggc gctaaattgt tcagtctaga
      961 tttggccgca ttaaccgcag gtgctaaatg gaaaggtgat ttcgaagaaa gattcaaagg
     1021 tgttttgaag gaaatcgaag aatcaaagac tctaattgtg ttattcattg atgaaattca
     1081 catgttaatg ggtaatggta aggacgacgc tgctaacatc ttgaagccag ctttgtccag
     1141 aggccaattg aaggtcatcg gtgccaccac caataacgaa tatagatcta ttgtggaaaa
     1201 ggatggtgcc tttgaaagaa gattccagaa aattgaagtc gctgaaccaa gtgtgagaca
     1261 aacagtggcc atattgagag gtctgcaacc aaagtatgaa atacatcatg gtgtaaggat
     1321 tctggatagc gccttagtca ccgcggctca attagccaag cgttacttgc catatagaag
     1381 attgccagat tctgctttgg atttagttga tatttcttgt gctggtgtcg ccgtcgcaag
     1441 agattctaag ccagaagaat tggattccaa ggaacgtcaa ttgcaattga ttcaagtaga
     1501 gataaaagct ctagagagag atgaagatgc cgactccacc actaaagata gattaaagtt
     1561 agctaggcag aaggaagctt cattgcaaga agaattggaa cctctaagac aacgttacaa
     1621 tgaagaaaag catggccatg aagaattgac acaagctaaa aagaaattgg atgaactgga
     1681 aaacaaggcc cttgatgctg aacgtagata tgatactgct accgccgctg atttaaggta
     1741 cttcgccatc ccagatatca aaaagcaaat cgaaaagctt gaagatcagg ttgctgagga
     1801 agagagacgt gctggtgcca actccatgat ccaaaatgtg gtcgactcag acaccatttc
     1861 tgaaacagct gcaagattga ctggtatccc tgttaagaag ttgtcagaat ctgaaaatga
     1921 aaaattgatt catatggaac gtgacttatc atctgaagtc gtgggccaaa tggatgccat
     1981 taaagctgtt tccaatgccg ttagattgtc tagatcaggt ttagctaatc caaggcaacc
     2041 agcatccttc ttatttttag gtttgtccgg ttccggtaaa actgaattgg ctaaaaaagt
     2101 tgctggattt ttgtttaatg atgaggacat gatgatcagg gtcgattgtt ctgaattaag
     2161 cgagaagtat gcggtctcta agttgttggg taccacggca ggttatgtcg ggtacgatga
     2221 aggtggcttt ttaactaacc aactgcaata caaaccatac tccgttttgt tattcgatga
     2281 agtagaaaag gcacatcctg atgttttgac tgtcatgcta caaatgttgg atgacggtag
     2341 aattactagt ggtcaaggta agacgatcga ctgttccaat tgtattgtca tcatgacttc
     2401 caatctaggt gctgaattta tcaattctca acaaggatca aagatccaag aatctaccaa
     2461 gaatttggtc atgggtgctg ttaggcaaca tttcagacca gaatttttga acagaatttc
     2521 tagtatagtc attttcaaca agctatctag aaaagctatt cataagatcg tggatattcg
     2581 tttgaaggaa attgaagaga gattcgagca aaatgataaa cattacaagt tgaatttaac
     2641 tcaagaggcc aaggacttct tggccaaata tggttattcc gatgatatgg gtgcacgtcc
     2701 actgaacagg ttaattcaaa acgaaatttt gaacaaactg gcactaagga tcttaaagaa
     2761 tgaaatcaag gataaggaaa ctgtcaatgt cgtcttgaag aagggtaaat ctcgtgatga
     2821 aaatgttcct gaggaagctg aagaatgtct ggaagttcta ccaaatcacg aagctactat
     2881 aggggctgac acgttaggtg atgacgataa tgaggacagt atggaaattg atgatgacct
     2941 agattaattt aatatagtgt gatttttaaa aactttcgaa caagaatcag taatataata
     3001 tatataatta ataaaactaa tggaatttgt ttaattgaac ttgacacccg agcaaaagag
     3061 ctcggggccc tagctaactg atccggctgc taacaaagcc caaaggaagc tgagttggct
     3121 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     3181 ggttttttgc tgaaaggagg aactatagat aattcttctt gaagacgaaa gggcctcgtg
     3241 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     3301 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3361 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3421 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3481 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3541 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3601 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3661 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3721 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3781 ttatgcagtg ctgccataag catgagtgat aacactgcgg ccaacttact tctgacaacg
     3841 atcggaggac cgaaggagct aaccgctttt tttcacaaca tgggggatca tgtaactcgc
     3901 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3961 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4021 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4081 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4141 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4201 tacacgacgg gcagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4261 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4321 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4381 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4441 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4501 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4561 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     4621 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4681 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4741 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4801 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagcattg agaaagcgcc
     4861 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4921 gagcgcacga gggagcttcc aggggggaac gcctggtatc tttatagtcc tgtcgggttt
     4981 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcc gagcctatgg
     5041 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5101 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5161 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5221 gaagt
//
