LOCUS       sequence-152938-        7471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Methionine ABC Transporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7471)
  AUTHORS   Kadaba NS, Kaiser JT, Johnson E, Lee A, Rees DC
  TITLE     The high-affinity E. coli methionine ABC transporter: structure and 
            allosteric regulation.
  JOURNAL   Science. 2008 Jul 11;321(5886):250-3. doi: 10.1126/science.1157987.
  PUBMED    18621668
REFERENCE   2  (bases 1 to 7471)
  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"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2008-07-11- 11"; volume: "321"; 
            issue: "5886"; pages: "250-3"
COMMENT     Sequence Label: MetNI (Lm)
FEATURES             Location/Qualifiers
     source          1..7471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(324..1406)
                     /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     1368..1387
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1407..1484)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        1600..1618
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1619..1643
                     /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             1658..1680
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2345..2362
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2429..2476
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2518..2536
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2537..2561
                     /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             2576..2598
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2611..2640
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2656..2670
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     terminator      3775..3822
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3997..4002,4003..4019,4020..4025))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(4052..4070)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4138..4242
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4243..4311,4312..5103)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4461..4480)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5274..5862
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5763..5782
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6016..6033
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6048..6190
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6116..6135)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6276..6298
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6292..6483)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
       61 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
      121 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
      181 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
      241 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
      301 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
      361 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
      421 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
      481 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
      541 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
      601 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
      661 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
      721 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
      781 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
      841 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
      901 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
      961 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     1021 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     1081 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     1141 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     1201 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     1261 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     1321 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     1381 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     1441 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     1501 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     1561 cgccgccgca aggaatggtg catgccgatc ccgcgaaatt aatacgactc actatagggg
     1621 aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga
     1681 tatacatatg tctgagccga tgatgtggct gctggttcgt ggcgtatggg aaacgctggc
     1741 aatgaccttc gtatccggtt tttttggctt tgtgattggt ctgccggttg gcgttctgct
     1801 ttatgtcacg cgtccggggc aaattattgc taacgcgaag ctgtatcgta ccgtttctgc
     1861 gattgtgaac attttccgtt ccatcccgtt cattatcttg cttgtatgga tgattccgtt
     1921 tacccgcgtt attgtcggta catcgattgg tttgcaggca gcgattgttc cgttaaccgt
     1981 tggtgcagca ccgtttattg cccgtatggt cgagaacgct ctgctggaga tcccaaccgg
     2041 gttaattgaa gcttcccgcg caatgggtgc cacgccgatg cagatcgtcc gtaaggtgct
     2101 gttaccggaa gcgctgccgg gtctggtgaa tgcggcaact atcaccctga ttaccctggt
     2161 cggttattcc gcgatgggtg gtgcagtcgg tgccggtggt ttaggtcaga ttggctatca
     2221 gtatggctac atcggctata acgcgacggt gatgaatacg gtactggtat tgctggtcat
     2281 tctggtttat ttaattcagt tcgcaggcga ccgcatcgtc cgggctgtca ctcgcaagtg
     2341 agagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga
     2401 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     2461 cttgaggggt tttttgctga aaggaggaac tatatccaga tctcgatccc gcgaaattaa
     2521 tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt
     2581 ttaactttaa gaaggagata taccatgggc catcatcatc atcatcatca tcatcatcac
     2641 agcagcggcc atatcgacga cgacgacaag catatgataa aactttcgaa tatcaccaaa
     2701 gtgttccacc agggcacccg caccatccag gcgttgaaca acgtcagcct gcatgtgcca
     2761 gctggacaaa tttatggcgt tatcggtgcc tcaggcgcgg gtaagagtac gcttatacgt
     2821 tgtgtaaacc tgctggagcg cccaaccgag ggtagcgtgc tggtcgatgg ccaggaactg
     2881 accacgctgt cagaatccga gttgaccaaa gctcgccgcc agattggtat gattttccag
     2941 cattttaacc tgctctcttc gcgtactgtt tttggcaacg tggctctgcc gctggagctg
     3001 gacaacacac cgaaagacga ggtcaaacgt cgcgtgacgg aattgctgtc attggttggt
     3061 cttggcgata agcatgatag ctacccgtcg aatctttccg gtgggcagaa acaacgtgtg
     3121 gcaattgccc gtgcgttagc cagcaatccc aaagtattgc tgtgtgatga agccaccagc
     3181 gcgctggacc cggcaacgac acgttctatt ctcgaactgc tgaaagacat caaccgccgt
     3241 ctggggttga cgattctgtt gatcacccac gaaatggacg ttgtgaagcg catttgtgat
     3301 tgcgtggcgg tcatcagcaa tggagaactg atcgagcagg acacggtaag tgaagtgttc
     3361 tcgcatccga aaacgccgct ggcgcagaag tttattcagt cgaccctgca tctggatatc
     3421 ccggaagatt accaggaacg tctgcaagcg gagccattta ctgactgcgt gccgatgctg
     3481 cgtctggagt ttaccggtca atcggtcgat gccccactgc tttctgaaac cgcgcgtcgt
     3541 ttcaacgtca acaacaacat tattagcgcg cagatggatt acgccggtgg cgttaagttc
     3601 ggcatcatgc tgactgaaat gcacggcaca caacaagata cgcaagccgc cattgcctgg
     3661 ctgcaggaac accatgtaaa agtagaggta ctgggttatg tctgactcga ggatccggct
     3721 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     3781 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     3841 tccggatatc ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat
     3901 ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct
     3961 gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct
     4021 gtcaaacatg agaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4081 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4141 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4201 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4261 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4321 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4381 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4441 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     4501 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4561 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4621 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4681 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4741 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     4801 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4861 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4921 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4981 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5041 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5101 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5161 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5221 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5281 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5341 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5401 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     5461 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5521 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5581 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5641 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5701 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5761 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5821 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5881 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5941 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     6001 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     6061 tttctcctta cgcatctgtg cggtatttca caccgcatat atggtgcact ctcagtacaa
     6121 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     6181 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     6241 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     6301 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     6361 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     6421 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     6481 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
     6541 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
     6601 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
     6661 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
     6721 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga
     6781 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
     6841 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
     6901 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gccaggaccc
     6961 aacgctgccc gagatgcgcc gcgtgcggct gctggagatg gcggacgcga tggatatgtt
     7021 ctgccaaggg ttggtttgcg cattcacagt tctccgcaag aattgattgg ctccaattct
     7081 tggagtggtg aatccgttag cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg
     7141 ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt atagggcggc gcctacaatc
     7201 catgccaacc cgttccatgt gctcgccgag gcggcataaa tcgccgtgac gatcagcggt
     7261 ccagtgatcg aagttaggct ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg
     7321 tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa
     7381 gcgagaagaa tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc cagcaagacg
     7441 tagcccagcg cgtcggccgc catgccggcg a
//
