LOCUS       sequence-223946-        5713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Wild Type Methionine ABC Transporter MetNI in pBAD Vector for 
            Transport Activity Assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5713)
  AUTHORS   Nguyen PT, Lai JY, Lee AT, Kaiser JT, Rees DC
  TITLE     Noncanonical role for the binding protein in substrate uptake by the
            MetNI methionine ATP Binding Cassette (ABC) transporter.
  JOURNAL   Proc Natl Acad Sci U S A. 2018 Oct 23. pii: 1811003115. doi: 
            10.1073/pnas.1811003115.
  PUBMED    30352853
REFERENCE   2  (bases 1 to 5713)
  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: "Proc Natl 
            Acad Sci U S A. 2018 Oct 23. pii: 1811003115. doi: 
            10.1073/pnas.1811003115."
COMMENT     Sequence Label: WT MetNI in pBAD
FEATURES             Location/Qualifiers
     source          1..5713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             334..363
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             379..393
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     primer_bind     complement(2114..2131)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2284..2370
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2462..2489
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2508..2599
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2600..2668,2669..3460)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2818..2837)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3631..4219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4120..4139
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4373..4390
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4405..4545
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4471..4490)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4631..4653
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4809..5687)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccga attcttaatg ggccatcatc atcatcatca tcatcatcat
      361 cacagcagcg gccatatcga cgacgacgac aagcatatga taaaactttc gaatatcacc
      421 aaagtgttcc accagggcac ccgcaccatc caggcgttga acaacgtcag cctgcatgtg
      481 ccagctggac aaatttatgg cgttatcggt gcctcaggcg cgggtaagag tacgcttata
      541 cgttgtgtaa acctgctgga gcgcccaacc gagggtagcg tgctggtcga tggccaggaa
      601 ctgaccacgc tgtcagaatc cgagttgacc aaagctcgcc gccagattgg tatgattttc
      661 cagcatttta acctgctctc ttcgcgtact gtttttggca acgtggctct gccgctggag
      721 ctggacaaca caccgaaaga cgaggtcaaa cgtcgcgtga cggaattgct gtcattggtt
      781 ggtcttggcg ataagcatga tagctacccg tcgaatcttt ccggtgggca gaaacaacgt
      841 gtggcaattg cccgtgcgtt agccagcaat cccaaagtat tgctgtgtga tgaagccacc
      901 agcgcgctgg acccggcaac gacacgttct attctcgaac tgctgaaaga catcaaccgc
      961 cgtctggggt tgacgattct gttgatcacc cacgaaatgg acgttgtgaa gcgcatttgt
     1021 gattgcgtgg cggtcatcag caatggagaa ctgatcgagc aggacacggt aagtgaagtg
     1081 ttctcgcatc cgaaaacgcc gctggcgcag aagtttattc agtcgaccct gcatctggat
     1141 atcccggaag attaccagga acgtctgcaa gcggagccat ttactgactg cgtgccgatg
     1201 ctgcgtctgg agtttaccgg tcaatcggtc gatgccccac tgctttctga aaccgcgcgt
     1261 cgtttcaacg tcaacaacaa cattattagc gcgcagatgg attacgccgg tggcgttaag
     1321 ttcggcatca tgctgactga aatgcacggc acacaacaag atacgcaagc cgccattgcc
     1381 tggctgcagg aacaccatgt aaaagtagag gtactgggtt atgtctgagc cgatgatgtg
     1441 gctgctggtt cgtggcgtat gggaaacgct ggcaatgacc ttcgtatccg gtttttttgg
     1501 ctttgtgatt ggtctgccgg ttggcgttct gctttatgtc acgcgtccgg ggcaaattat
     1561 tgctaacgcg aagctgtatc gtaccgtttc tgcgattgtg aacattttcc gttccatccc
     1621 gttcattatc ttgcttgtat ggatgattcc gtttacccgc gttattgtcg gtacatcgat
     1681 tggtttgcag gcagcgattg ttccgttaac cgttggtgca gcaccgttta ttgcccgtat
     1741 ggtcgagaac gctctgctgg agatcccaac cgggttaatt gaagcttccc gcgcaatggg
     1801 tgccacgccg atgcagatcg tccgtaaggt gctgttaccg gaagcgctgc cgggtctggt
     1861 gaatgcggca actatcaccc tgattaccct ggtcggttat tccgcgatgg gtggtgcagt
     1921 cggtgccggt ggtttaggtc agattggcta tcagtatggc tacatcggct ataacgcgac
     1981 ggtgatgaat acggtactgg tattgctggt cattctggtt tatttaattc agttcgcagg
     2041 cgaccgcatc gtccgggctg tcactcgcaa gtaagaagct tggctgtttt ggcggatgag
     2101 agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga
     2161 atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga
     2221 aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg
     2281 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     2341 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag
     2401 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag
     2461 cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt
     2521 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2581 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2641 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2701 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     2761 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     2821 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     2881 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     2941 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3001 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3061 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3121 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3181 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3241 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3301 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3361 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3421 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3481 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3541 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3601 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3661 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3721 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     3781 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3841 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3901 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3961 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4021 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4081 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4141 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4201 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4261 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4321 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4381 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4441 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4501 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     4561 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4621 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4681 aggcagcaga tcaattcgcg cgcgaaggcg aagcggcatg cataatgtgc ctgtcaaatg
     4741 gacgaagcag ggattctgca aaccctatgc tactccgtca agccgtcaat tgtctgattc
     4801 gttaccaatt atgacaactt gacggctaca tcattcactt tttcttcaca accggcacgg
     4861 aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaaata gagttgatcg
     4921 tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct caaaagcagc
     4981 ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc taactgctgg
     5041 cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac gctggcgata
     5101 tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg tacccgatta
     5161 tccatcggtg gatggagcga ctcgttaatc gcttccatgc gccgcagtaa caattgctca
     5221 agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc gttaatgatt
     5281 tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta
     5341 ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa
     5401 acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg aatctctcct
     5461 ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg atttttcacc
     5521 accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag cggtcggtcg
     5581 ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac cagatgggca
     5641 ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact tttcatactc
     5701 ccgccattca gag
//
