LOCUS       sequence-148135-        4035 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 4035)
  AUTHORS   Martin A, Baker TA, Sauer RT
  TITLE     Rebuilt AAA + motors reveal operating principles for ATP-fuelled 
            machines.
  JOURNAL   Nature. 2005 Oct 20. 437(7062):1115-20.
  PUBMED    16237435
REFERENCE   2  (bases 1 to 4035)
  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: "Nature. 
            2005 Oct 20. 437(7062):1115-20."
COMMENT     Sequence Label: ClpP-his
FEATURES             Location/Qualifiers
     source          1..4035
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..15,16..21,22..38,39..45)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    21..37
                     /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)."
     protein_bind    53..69
                     /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             88..99
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             733..750
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      773..867
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             911..1570
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(977..996)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1635..1721
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1768..1790)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1876..2016
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1931..1950
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2031..2048)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2202..2790)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2282..2301)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2961..3752,3753..3821))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3584..3603
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3822..3926)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3994..4012
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taatagattc aattgtgagc
       61 ggataacaat ttcacacaga attcattaaa gaggagaaat taagcatgcc atacagcggc
      121 gaacgagata actttgcacc ccatatggcg ctggtgccga tggtcattga acagacctca
      181 cgcggtgagc gctcttttga tatctattct cgtctactta aggaacgcgt catttttctg
      241 actggccagg ttgaagacca catggctaac ctgattgtgg cgcagatgct gttcctggaa
      301 gcggaaaacc cagaaaaaga tatctatctg tacattaact ccccaggcgg ggtgatcact
      361 gccgggatgt ctatctatga caccatgcag tttatcaagc ctgatgtcag caccatctgt
      421 atgggccagg cggcctcgat gggcgctttc ttgctgaccg caggggcaaa aggtaaacgt
      481 ttttgcctgc cgaattcgcg cgtgatgatt caccaaccgt tgggcggcta ccagggccag
      541 gcgaccgata tcgaaattca tgcccgtgaa attctgaaag ttaaagggcg catgaatgaa
      601 cttatggcgc ttcatacggg tcaatcatta gaacagattg aacgtgatac cgagcgcgat
      661 cgcttccttt ccgcccctga agcggtggaa tacggtctgg tcgattcgat tctgacccat
      721 cgtaatagat ctcatcacca tcaccatcac taagcttaat tagctgagct tggactcctg
      781 ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc
      841 cgccgggcgt tttttattgg tgagaatcca agctagcttg gcgagatttt caggagctaa
      901 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
      961 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
     1021 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
     1081 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaatttc gtatggcaat
     1141 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga
     1201 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
     1261 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
     1321 gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga
     1381 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta
     1441 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     1501 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg
     1561 cggggcgtaa tttttttaag gcagttattg gtgcccttaa acgcctgggg taatgactct
     1621 ctagcttgag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     1681 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccctct agagctgcct
     1741 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     1801 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     1861 tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg
     1921 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     1981 ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact
     2041 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2101 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2161 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2221 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2281 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2341 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2401 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2461 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2521 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2581 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2641 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2701 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2761 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2821 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2881 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2941 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3001 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3061 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3121 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3181 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3241 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3301 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3361 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3421 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3481 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3541 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3601 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3661 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3721 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3781 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3841 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3901 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     3961 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     4021 cttcacctcg agaaa
//
