LOCUS       sequence-232032-        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"
     primer_bind     29..47
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(71..85,86..91,92..108,109..115)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    91..107
                     /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    123..139
                     /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             158..169
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1194..1288
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1332..1991
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1398..1417)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2056..2142
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2235..3317)
                     /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
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAISVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     3279..3298
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3318..3395)
                     /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(3479..3501)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3587..3727
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3642..3661
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3742..3759)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3913..4501)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3993..4012)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4672..5463,5464..5532))
                     /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     5295..5314
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5533..5637)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
       61 cctcgagaaa tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taatagattc
      121 aattgtgagc ggataacaat ttcacacaga attcattaaa gaggagaaat taactatgaa
      181 taagaaggtg ttaaccctgt ctgctgtgat ggccagcatg ttattcggtg ccgctgcaca
      241 cgctgctgat actcgcattg gtgtaacaat ctataagtac gacgataact ttatgtctgt
      301 agtgcgcaag gctattgagc aagatgcgaa agccgcgcca gatgttcagc tgctgatgaa
      361 tgattctcag aatgaccagt ccaagcagaa cgatcagatc gacgtattgc tggcgaaagg
      421 ggtgaaggca ctggcaatca acctggttga cccggcagct gcgggtacgg tgattgagaa
      481 agcgcgtggg caaaacgtgc cggtggtttt cttcaacaaa gaaccgtctc gtaaggcgct
      541 ggatagctac gacaaagcct actacgttgg cactgactcc aaagagtccg gcattattca
      601 aggcgatttg attgctaaac actgggcggc gaatcagggt tgggatctga acaaagacgg
      661 tcagattcag ttcgtactgc tgaaaggtga accgggccat ccggatgcag aagcacgtac
      721 cacttacgtg attaaagaat tgaacgataa aggcatcaaa actgaacagt tacagttaga
      781 taccgcaatg tgggacaccg ctcaggcgaa agataagatg gacgcctggc tgtctggccc
      841 gaacgccaac aaaatcgaag tggttatcgc caacaacgat gcgatggcaa tgggcgcggt
      901 tgaagcgctg aaagcacaca acaagtccag cattccggtg tttggcgtcg atgcgctgcc
      961 agaagcgctg gcgtgtgtga aatccggtgc actggcgggc accgtactga acgatgctaa
     1021 caaccaggcg aaagcgacct ttgatctggc gaaaaacctg gccgatggta aaggtgcggc
     1081 tgatggcacc aactggaaaa tctgcaacaa agtggtccgc gtaccttatg ttggcgtaga
     1141 taaagacaac ctggctgaat tcagcaagaa ataagcttaa ttagctgagc ttggactcct
     1201 gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg
     1261 ccgccgggcg ttttttattg gtgagaatcc aagctagctt ggcgagattt tcaggagcta
     1321 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
     1381 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
     1441 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
     1501 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattt cgtatggcaa
     1561 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
     1621 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
     1681 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
     1741 ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg
     1801 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt
     1861 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
     1921 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
     1981 gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggg gtaatgactc
     2041 tctagcttga ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     2101 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccctc tagattacgt
     2161 gcagtcgatg ataagctgtc aaacatgaga attgtgccta atgagtgagc taacttacat
     2221 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     2281 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     2341 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     2401 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     2461 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc
     2521 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     2581 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
     2641 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     2701 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     2761 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     2821 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     2881 aacgccggaa cattagtgca ggcagcttcc acagaaatgg catcctggtc atccagcgga
     2941 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     3001 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     3061 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     3121 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     3181 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     3241 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     3301 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     3361 ataccgcgaa aggttttgca ccattcgatg gtgtcggaat ttcgggcagc gttgggtcct
     3421 ggccacgggt gcgcatgatc tagagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     3481 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     3541 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     3601 agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt
     3661 actgagagtg caccacatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     3721 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3781 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3841 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3901 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3961 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4021 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4081 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4141 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4201 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4261 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4321 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     4381 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4441 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4501 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4561 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4621 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4681 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4741 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4801 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4861 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4921 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4981 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5041 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5101 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5161 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5221 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5281 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5341 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5401 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5461 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5521 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5581 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5641 atttccccga aaagtgccac ctgacgtcta agaaac
//
