LOCUS       sequence-227052-        5676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  precusor D-galactose/methyl-galactoside ABC transporter periplasmic 
            binding protein, mglB, L244C, D287C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5676)
  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 5676)
  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: pAL725
FEATURES             Location/Qualifiers
     source          1..5676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1019..1113
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1157..1816
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1223..1242)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1881..1967
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2060..3142)
                     /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     3104..3123
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3143..3220)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(3304..3326)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3412..3552
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3467..3486
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3567..3584)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3738..4326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3818..3837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4497..5288,5289..5357))
                     /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     5120..5139
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5358..5462)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5530..5548
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(5572..5586,5587..5592,5593..5609,5610..5616)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    5592..5608
                     /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    5624..5640
                     /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             5659..5670
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgaataaga aggtgttaac cctgtctgct gtgatggcca gcatgttatt cggtgccgct
       61 gcacacgctg ctgatactcg cattggtgta acaatctata agtacgacga taactttatg
      121 tctgtagtgc gcaaggctat tgagcaagat gcgaaagccg cgccagatgt tcagctgctg
      181 atgaatgatt ctcagaatga ccagtccaag cagaacgatc agatcgacgt attgctggcg
      241 aaaggggtga aggcactggc aatcaacctg gttgacccgg cagctgcggg tacggtgatt
      301 gagaaagcgc gtgggcaaaa cgtgccggtg gttttcttca acaaagaacc gtctcgtaag
      361 gcgctggata gctacgacaa agcctactac gttggcactg actccaaaga gtccggcatt
      421 attcaaggcg atttgattgc taaacactgg gcggcgaatc agggttggga tctgaacaaa
      481 gacggtcaga ttcagttcgt actgctgaaa ggtgaaccgg gccatccgga tgcagaagca
      541 cgtaccactt acgtgattaa agaattgaac gataaaggca tcaaaactga acagttacag
      601 ttagataccg caatgtggga caccgctcag gcgaaagata agatggacgc ctggctgtct
      661 ggcccgaacg ccaacaaaat cgaagtggtt atcgccaaca acgatgcgat ggcaatgggc
      721 gcggttgaag cgctgaaagc acacaacaag tccagcattc cggtgtttgg cgtcgatgcg
      781 ctgccagaag cgctggcgtg tgtgaaatcc ggtgcactgg cgggcaccgt actgaacgat
      841 gctaacaacc aggcgaaagc gacctttgat ctggcgaaaa acctggccga tggtaaaggt
      901 gcggctgatg gcaccaactg gaaaatctgc aacaaagtgg tccgcgtacc ttatgttggc
      961 gtagataaag acaacctggc tgaattcagc aagaaataag cttaattagc tgagcttgga
     1021 ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc
     1081 ggttgccgcc gggcgttttt tattggtgag aatccaagct agcttggcga gattttcagg
     1141 agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca
     1201 atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca
     1261 gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt
     1321 ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg aatttcgtat
     1381 ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt
     1441 ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca
     1501 gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc
     1561 taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag
     1621 ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa
     1681 atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt
     1741 ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg
     1801 gcagggcggg gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc ctggggtaat
     1861 gactctctag cttgaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     1921 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccctctagat
     1981 tacgtgcagt cgatgataag ctgtcaaaca tgagaattgt gcctaatgag tgagctaact
     2041 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2101 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     2161 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     2221 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     2281 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     2341 tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     2401 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     2461 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     2521 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     2581 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     2641 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     2701 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     2761 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     2821 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     2881 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     2941 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     3001 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     3061 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     3121 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     3181 atgccatacc gcgaaaggtt ttgcaccatt cgatggtgtc ggaatttcgg gcagcgttgg
     3241 gtcctggcca cgggtgcgca tgatctagag ctgcctcgcg cgtttcggtg atgacggtga
     3301 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     3361 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     3421 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
     3481 attgtactga gagtgcacca catgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     3541 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3601 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3661 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3721 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3781 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3841 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3901 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3961 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4021 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4081 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4141 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     4201 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4261 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4321 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4381 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4441 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4501 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4561 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4621 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4681 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4741 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4801 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4861 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4921 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4981 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5041 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5101 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5161 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5221 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5281 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5341 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5401 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5461 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     5521 acctataaaa ataggcgtat cacgaggccc tttcgtcttc acctcgagaa atcataaaaa
     5581 atttatttgc tttgtgagcg gataacaatt ataatagatt caattgtgag cggataacaa
     5641 tttcacacag aattcattaa agaggagaaa ttaact
//
