LOCUS       sequence-402649-        5247 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 5247)
  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 5247)
  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 Y819W
FEATURES             Location/Qualifiers
     source          1..5247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    10..34
                     /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             49..71
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             84..113
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             126..137
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3055..3073)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3069..3116
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3161..3179)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3247..3351
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3352..3420,3421..4212)
                     /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(3570..3589)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4383..4971
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4872..4891
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5125..5142
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     join(5238..5247,1..10)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5238..5247,1..9)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
       61 taagaaggag atataccatg ggccatcatc atcatcatca tcatcatcat cacagcagcg
      121 gccatatcga aggtcgtgga tccgggatga acgaccaaac gcaatttaca gaaagggctc
      181 taacgatttt gacgttggct caaaaattgg cttcggatca tcaacatcca caattacaac
      241 ctatacatat tctagctgcc ttcattgaaa cgccagaaga tggatcagtc ccttacctac
      301 agaatctaat tgagaagggc cgttacgact atgatctttt caagaaagtg gttaatagaa
      361 atctagtaag aattcctcaa cagcaacctg cacctgcgga gataactcca agttatgctt
      421 tggggaaagt ccttcaagac gctgctaaga ttcaaaaaca acagaaggac tcatttatag
      481 cgcaagacca tatattgttt gctctattca atgattcgtc tattcagcaa atatttaagg
      541 aagctcaagt agatattgag gccatcaagc aacaagctct tgaacttcgt ggtaacacta
      601 gaattgactc tcgtggcgct gatacgaaca cacctttgga atatttatca aagtacgcca
      661 ttgatatgac tgagcaggct cgtcaaggta aacttgaccc tgtcatcggc cgtgaagaag
      721 aaataagaag cactattaga gttttagcaa gaagaattaa gtccaaccca tgtttaattg
      781 gtgagccagg tatcggtaag accgctatta ttgaaggtgt tgctcaaaga atcattgacg
      841 atgacgttcc cactatctta caaggcgcta aattgttcag tctagatttg gccgcattaa
      901 ccgcaggtgc taaatacaaa ggtgatttcg aagaaagatt caaaggtgtt ttgaaggaaa
      961 tcgaagaatc aaagactcta attgtgttat tcattgatga aattcacatg ttaatgggta
     1021 atggtaagga cgacgctgct aacatcttga agccagcttt gtccagaggc caattgaagg
     1081 tcatcggtgc caccaccaat aacgaatata gatctattgt ggaaaaggat ggtgcctttg
     1141 aaagaagatt ccagaaaatt gaagtcgctg aaccaagtgt gagacaaaca gtggccatat
     1201 tgagaggtct gcaaccaaag tatgaaatac atcatggtgt aaggattctg gatagcgcct
     1261 tagtcaccgc ggctcaatta gccaagcgtt acttgccata tagaagattg ccagattctg
     1321 ctttggattt agttgatatt tcttgtgctg gtgtcgccgt cgcaagagat tctaagccag
     1381 aagaattgga ttccaaggaa cgtcaattgc aattgattca agtagagata aaagctctag
     1441 agagagatga agatgccgac tccaccacta aagatagatt aaagttagct aggcagaagg
     1501 aagcttcatt gcaagaagaa ttggaacctc taagacaacg ttacaatgaa gaaaagcatg
     1561 gccatgaaga attgacacaa gctaaaaaga aattggatga actggaaaac aaggcccttg
     1621 atgctgaacg tagatatgat actgctaccg ccgctgattt aaggtacttc gccatcccag
     1681 atatcaaaaa gcaaatcgaa aagcttgaag atcaggttgc tgaggaagag agacgtgctg
     1741 gtgccaactc catgatccaa aatgtggtcg actcagacac catttctgaa acagctgcaa
     1801 gattgactgg tatccctgtt aagaagttgt cagaatctga aaatgaaaaa ttgattcata
     1861 tggaacgtga cttatcatct gaagtcgtgg gccaaatgga tgccattaaa gctgtttcca
     1921 atgccgttag attgtctaga tcaggtttag ctaatccaag gcaaccagca tccttcttat
     1981 ttttaggttt gtccggttcc ggtaaaactg aattggctaa aaaagttgct ggatttttgt
     2041 ttaatgatga ggacatgatg atcagggtcg attgttctga attaagcgag aagtatgcgg
     2101 tctctaagtt gttgggtacc acggcaggtt atgtcgggta cgatgaaggt ggctttttaa
     2161 ctaaccaact gcaatacaaa ccatactccg ttttgttatt cgatgaagta gaaaaggcac
     2221 atcctgatgt tttgactgtc atgctacaaa tgttggatga cggtagaatt actagtggtc
     2281 aaggtaagac gatcgactgt tccaattgta ttgtcatcat gacttccaat ctaggtgctg
     2341 aatttatcaa ttctcaacaa ggatcaaaga tccaagaatc taccaagaat ttggtcatgg
     2401 gtgctgttag gcaacatttc agaccagaat ttttgaacag aatttctagt atagtcattt
     2461 tcaacaagct atctagaaaa gctattcata agatcgtgga tattcgtttg aaggaaattg
     2521 aagagagatt cgagcaaaat gataaacatt acaagttgaa tttaactcaa gaggccaagg
     2581 acttcttggc caaatatggt tggtccgatg atatgggtgc acgtccactg aacaggttaa
     2641 ttcaaaacga aattttgaac aaactggcac taaggatctt aaagaatgaa atcaaggata
     2701 aggaaactgt caatgtcgtc ttgaagaagg gtaaatctcg tgatgaaaat gttcctgagg
     2761 aagctgaaga atgtctggaa gttctaccaa atcacgaagc tactataggg gctgacacgt
     2821 taggtgatga cgataatgag gacagtatgg aaattgatga tgacctagat taatttaata
     2881 tagtgtgatt tttaaaaact ttcgaacaag aatcagtaat ataatatata taattaataa
     2941 aactaatgga atttgtttaa ttgaacttga cacccgagca aaagagctcg gggccctagc
     3001 taactgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
     3061 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
     3121 aaggaggaac tatatccgga taattcttga agacgaaagg gcctcgtgat acgcctattt
     3181 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3241 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3301 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3361 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3421 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3481 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3541 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     3601 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3661 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3721 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3781 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3841 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3901 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3961 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4021 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4081 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4141 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4201 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4261 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4321 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4381 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4441 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4501 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4561 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4621 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4681 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4741 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4801 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4861 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4921 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4981 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5041 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5101 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccag
     5161 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     5221 tctcgatccc gcgaaattaa tacgact
//
