LOCUS       sequence-232465-        6533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  precursor slow-folding rbsB with the mutations Alanine 248 
            Threonine, Alanine 188 Cysteine and Glutamic acid 246 Cysteine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6533)
  AUTHORS   Bariya P, Randall LL
  TITLE     Co-assembly of SecYEG and SecA fully restores the properties of the 
            native translocon.
  JOURNAL   J Bacteriol. 2018 Oct 1. pii: JB.00493-18. doi: 10.1128/JB.00493-18.
  PUBMED    30275279
REFERENCE   2  (bases 1 to 6533)
  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: "J 
            Bacteriol. 2018 Oct 1. pii: JB.00493-18. doi: 10.1128/JB.00493-18."
COMMENT     Sequence Label: pAL942
FEATURES             Location/Qualifiers
     source          1..6533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     571..590
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(610..687)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     893..912
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     996..1015
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        996..1014
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1015..1039
                     /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             1054..1076
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2025..2043)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2039..2086
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2320..2338)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2406..2510
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2511..2579,2580..3371)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRMHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2729..2748)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3542..4130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4031..4050
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4284..4301
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4316..4455
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4381..4400)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4541..4563
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4557..4748)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6060..6533,1..609))
                     /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
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSVRLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 tttatgccag ccagccagac gcagacgcac cgagacagaa cttaatgggc ccgctaacag
       61 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
      121 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
      181 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
      241 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
      301 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
      361 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
      421 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
      481 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
      541 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
      601 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
      661 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct
      721 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
      781 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
      841 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
      901 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
      961 cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt
     1021 gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata
     1081 catatgaaca tgaaaaaact ggctaccctg gtttccgctg ttgcgctaag cgccaccgtc
     1141 agtgcgaatg cgatggcaaa agacaccatc gcgctggtgg tctccacgct taacaacccg
     1201 ttctttgtat cgctgaaaga tggcgcgcag aaagaggcgg ataaacttgg ctataacctg
     1261 gtggtgctgg actcccagaa caacccggcg aaagagctgg cgaacgtgca ggacttaacc
     1321 gttcgcggca caaaaattct gctgattaac ccgaccgact ccgacgcagt gggtaatgct
     1381 gtgaagatgg ctaaccaggc gaacatcccg gttatcactc ttgaccgcca ggcaacgaaa
     1441 ggtgaagtgg tgagccacat tgcttctgat aacgtactgg gcggcaaaat cgctggtgat
     1501 tacatcgcga agaaagcggg tgaaggtgcc aaagttatcg agctgcaagg cattgctggt
     1561 acatccgcag cccgtgaacg tggcgaaggc ttccagcagg ccgttgctgc tcacaagttt
     1621 aatgttcttg ccagccagcc agcagatttt gatcgcatta aaggtttgaa cgtaatgcag
     1681 aacctgttga ccgctcatcc ggatgttcag gctgtattct gtcagaatga tgaaatggcg
     1741 ctgggcgcgc tgcgcgcact gcaaactgcc ggtaaatcgg atgtgatggt cgtcggattt
     1801 gacggtacac cggatggcga aaaagcggtg aatgatggca aactagcagc gactatcgct
     1861 cagctacccg atcagattgg cgcgaaaggc gtctgtacca cagataaagt gctgaaaggc
     1921 gagaaagttc aggctaagta tccggttgat ctgaaactgg ttgttaagca gtagggatcc
     1981 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     2041 agcataaccc cttggggcct ctaaacgggt ctttaggggt tttttgctga aaggaggaac
     2101 tatatccgga tatcccgcaa gaggcccggc agtaccggca taaccaagcc tatgcctaca
     2161 gcatccaggg tgacggtgcc gaggatgacg atgagcgcat tgttagattt catacacggt
     2221 gcctgactgc gttagcaatt taactgtgat aaactaccgc attaaagcta gcttatcgat
     2281 gataagctgt caaacatgag aattcttgaa gacgaaaggg cctcgtgata cgcctatttt
     2341 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     2401 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2461 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2521 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2581 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2641 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2701 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     2761 ccgggcaaga gcaactcggt cgccgcatgc actattctca gaatgacttg gttgagtact
     2821 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2881 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2941 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3001 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     3061 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccgacaac
     3121 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3181 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3241 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3301 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3361 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3421 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3481 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3541 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3601 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3661 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     3721 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3781 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3841 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3901 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3961 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4021 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4081 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4141 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4201 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4261 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     4321 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     4381 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     4441 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     4501 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     4561 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     4621 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     4681 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     4741 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     4801 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     4861 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     4921 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     4981 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     5041 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     5101 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     5161 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag
     5221 gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat
     5281 atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca
     5341 attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg
     5401 cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta
     5461 caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca
     5521 gcggtccaat gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct
     5581 gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc
     5641 cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg aacgccagca
     5701 agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
     5761 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg
     5821 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc
     5881 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
     5941 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     6001 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     6061 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6121 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     6181 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     6241 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     6301 catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc
     6361 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     6421 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     6481 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag ata
//
