LOCUS       sequence-190588-        5791 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of BtuCD in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5791)
  AUTHORS   Locher KP, Lee AT, Rees DC
  TITLE     The E. coli BtuCD structure: a framework for ABC transporter 
            architecture and mechanism.
  JOURNAL   Science. 2002 May 10;296(5570):1091-8.
  PUBMED    12004122
REFERENCE   2  (bases 1 to 5791)
  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: "Science."; 
            date: "2002-05-10"; volume: "296(5570)"; pages: "1091-8"
COMMENT     Sequence Label: BtuCD
FEATURES             Location/Qualifiers
     source          1..5791
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     342..361
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     595..612
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    627..766
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(692..711)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     852..874
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(868..1059)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1868..2950)
                     /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     2912..2931
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2951..3028)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3234..3253
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3337..3356
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3337..3355
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3356..3380
                     /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             3395..3417
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3430..3459
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3475..3489
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     RBS             4486..4508
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5339..5357)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5353..5400
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5524..5543)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5734..5755
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
       61 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
      121 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
      181 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
      241 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
      301 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
      361 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
      421 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
      481 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
      541 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
      601 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
      661 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
      721 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
      781 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
      841 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
      901 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
      961 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     1021 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     1081 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     1141 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     1201 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     1261 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     1321 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     1381 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     1441 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     1501 caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc
     1561 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     1621 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     1681 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     1741 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     1801 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     1861 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1921 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     1981 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     2041 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     2101 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa
     2161 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     2221 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     2281 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     2341 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     2401 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     2461 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     2521 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     2581 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     2641 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     2701 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     2761 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     2821 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     2881 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
     2941 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     3001 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
     3061 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     3121 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     3181 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     3241 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     3301 ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat
     3361 tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat
     3421 accatgggcc atcatcatca tcatcatcat catcatcaca gcagcggcca tatcgacgac
     3481 gacgacaagc atatgctgac acttgcccgc caacaacagc gacaaaatat tcgctggtta
     3541 ttatgcctgt cagttttgat gctgctggcg cttctcttaa gcctttgcgc cggtgaacaa
     3601 tggatctcgc caggtgactg gtttactcct cgtggcgaac tgttcgtctg gcaaattcgc
     3661 ctgccacgta cgctggctgt attgctggtt ggtgcggcgc tggctatatc cggcgctgta
     3721 atgcaggcgt tgtttgaaaa tcctctggca gaacctggac tacttggcgt ctctaacggc
     3781 gcaggcgtgg ggcttatcgc cgcggtattg cttgggcaag ggcaactccc caactgggcg
     3841 ctagggctgt gtgcgattgc tggcgcgctt atcatcactt taatactctt acgtttcgcc
     3901 cgtcgtcatc tttcgaccag tcggttattg ctggctggcg ttgcattagg gattatctgt
     3961 agcgcactaa tgacgtgggc tatctacttt tccacctcag ttgatttgcg tcagctgatg
     4021 tactggatga tgggcggttt tggcggcgta gactggcggc aaagctggct gatgctggca
     4081 ttgatccccg tgttgttgtg gatctgttgt cagtccaggc cgatgaatat gttagcactt
     4141 ggcgagatct cggcgcggca actgggttta cccctgtggt tctggcgcaa tgtgctggtg
     4201 gcagcgaccg gctggatggt tggcgtcagt gtggcgctgg cgggtgctat cggctttatt
     4261 ggtctggtga tcccccatat tctccggttg tgtggtttaa ccgatcatcg cgtattactt
     4321 cccggctgcg cgctggcagg ggcgagcgca ttgctgctgg ccgatattgt agcgcgcctg
     4381 gcattagctg ccgcagagct gcctattggc gtggtcaccg caacgttagg tgcgccggtg
     4441 tttatctggt tattgttaaa agcaggacgt tagtctagaa ataattttgt ttaactttaa
     4501 gaaggagata tacatatgtc tattgtgatg cagttacaag atgttgcgga atctacccgc
     4561 ctggggccgc tttctggcga ggttcgggct ggggagatcc tgcacctggt ggggccgaat
     4621 ggcgcgggta agagtacctt actggcgcga atggccggaa tgaccagcgg taagggaagc
     4681 attcagttcg cggggcaacc actggaagca tggtccgcaa caaaactcgc gctgcatcgc
     4741 gcctatcttt cacaacagca gacgccgccg tttgcaacgc cggtctggca ctacctgaca
     4801 ctgcatcagc acgataaaac gcgtaccgaa ctactgaatg atgtcgcagg ggcgctggct
     4861 cttgatgaca aactcggacg tagcaccaat caactttccg gcggtgaatg gcaacgcgta
     4921 cgtcttgctg cggtggtgtt gcaaatcaca ccacaagcca atcccgcagg ccaattgctg
     4981 cttcttgatg agccgatgaa cagtcttgat gttgcgcaac aaagtgcgtt agacaaaatt
     5041 ctgagcgcgc tgtgtcagca aggactggcg attgtgatga gcagtcacga tctcaaccac
     5101 acattgcgtc atgcgcatcg ggcgtggttg ctaaaaggtg gaaaaatgct ggccagtgga
     5161 cgcagggaag aggtgctcac gccgccaaat ctggcgcagg cctatgggat gaattttcgc
     5221 cgtctggata tcgaaggtca cagaatgctg atttcgacca tcctcgagta gcaccaccac
     5281 caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     5341 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     5401 ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa
     5461 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     5521 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5581 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5641 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5701 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5761 cactcaaccc tatctcggtc tattcttttg a
//
