LOCUS       sequence-444316-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation of yebST knockout in E. coli K-12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7073)
  AUTHORS   Isom GL, Davies NJ, Chong ZS, Bryant JA, Jamshad M, Sharif M, 
            Cunningham AF, Knowles TJ, Chng SS, Cole JA, Henderson IR
  TITLE     MCE domain proteins: conserved inner membrane lipid-binding proteins
            required for outer membrane homeostasis.
  JOURNAL   Sci Rep. 2017 Aug 17;7(1):8608. doi: 10.1038/s41598-017-09111-6.
  PUBMED    28819315
REFERENCE   2  (bases 1 to 7073)
  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: "Sci Rep."; 
            date: "2017-08-17"; volume: "7(1):8608. doi"; pages: " 
            10.1038/s41598-017-09111-6"
COMMENT     Sequence Label: pET17b-yebST
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..131
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        112..130
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             162..184
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4142..4160)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4156..4203
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4248..4266)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4334..4438
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4439..4507,4508..5299)
                     /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(4657..4676)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5470..6058
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5959..5978
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6212..6229
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6244..6383
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6309..6328)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6469..6491
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6485..6676)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
       61 cgcacccgtg gccaggaccc aacgctgccc gagatctcga tcccgcgaaa ttaatacgac
      121 tcactatagg gagaccacaa cggtttccct ctagaaataa ttttgtttaa ctttaagaag
      181 gagatataca tatggctctt aacacaccac aaattacgcc gacaaaaaag ataacagtga
      241 gggcaatcgg cgaggaactg ccgcgtggtg attaccaacg ttgcccgcaa tgtgacatgc
      301 tgtttagcct gcccgagata aattctcatc aaagtgccta ttgtccgcgc tgtcaggcaa
      361 aaattcgtga cgggcgcgac tggtcgctaa cgcgcctggc ggcaatggcc ttcaccatgc
      421 tgttgcttat gccgtttgcc tggggcgaac cgctgttgca tatctggctg ttaggcattc
      481 gtattgacgc caacgttatg caaggcatct ggcaaatgac caaacagggc gatgcgataa
      541 cggggtcgat ggtctttttc tgcgttattg gtgccccgct cattctggtg acctccatag
      601 cttatttatg gtttggtaac cgactgggaa tgaatttacg tccggtactg ctgatgcttg
      661 agcgacttaa agagtgggta atgctcgata tctacctggt cggcattggc gttgcttcta
      721 taaaggtaca ggattatgcc catatccagg ctggtgtcgg cttgttctct tttgtggcgt
      781 tggtgatttt aacgacggtg acgttgtcac atcttaatgt cgaagaactg tgggagcgat
      841 tttatccgca gcgccccgct acgcgtaggg acgagaaact tcgtgtctgc ctagggtgcc
      901 attttaccgg ctatccagat cagcgtggtc gctgcccgcg ttgccatatc ccgctacgcc
      961 tgcgtcgccg tcatagtctg caaaaatgct gggcggcgct gttagcgtca atcgttttat
     1021 tgttacctgc caacctgttg cctatttcta tcatttatct gaacggagga cggcaggaag
     1081 atacaattct ttccggaatt atgtcgctgg caagtagcaa cattgcggtt gcaggaatcg
     1141 tgtttatcgc cagtattctg gtcccgttta ctaaagtgat cgtcatgttc actttactgt
     1201 tgagcattca ttttaaatgc cagcaaggtt tacgcacacg cattctgtta ctgcggatgg
     1261 tgacctggat tggtcgctgg tcgatgctcg acctgtttgt catatcttta accatgtcgc
     1321 tgattaatcg cgatcagatc ctcgctttta ctatgggacc ggctgcgttt tatttcggcg
     1381 cagcggtaat tttgactatt cttgctgtgg aatggctgga cagccgctta ctttgggatg
     1441 cacatgagtc aggaaacgcc cgcttcgacg actgaagcgc agattaaaaa taaacgccgt
     1501 atctcacctt tctggctgct gccgttcatc gcgctaatga ttgccagttg gctgatttgg
     1561 gacagttatc aggaccgggg taataccgtc accatcgact ttatgtcggc ggatggtatt
     1621 gttccaggcc gtacgcctgt tcgttatcag ggcgttgaag tcggaacagt gcaggacatc
     1681 agcctcagcg acgatcttcg taagattgaa gtcaaggtca gcatcaagtc cgatatgaaa
     1741 gatgcgctgc gcgaagagac tcagttctgg ctggtgacgc caaaagcatc gttggcaggt
     1801 gtctccgggc tggacgccct cgtcggtggg aactatatcg gcatgatgcc cgggaaaggt
     1861 aaagagcagg atcactttgt cgcactcgat acccaaccga aatatcggct ggacaatggc
     1921 gatctgatga tccacctgca agcccccgat ctcggttcgc tgaacagcgg ttcattggtc
     1981 tatttccgca agatcccggt gggaaaagtc tacgactatg ccatcaatcc caacaagcaa
     2041 ggcgtggtga ttgatgtcct gatcgagcgg cgttttaccg atctggtgaa aaaaggtagc
     2101 cgtttctgga acgtttccgg cgttgatgcc aacgtcagta tcagtggcgc gaaggtgaaa
     2161 ctggaaagct tggcggcact ggttaacggt gcgattgcct tcgattcacc agaagagtcg
     2221 aaacctgccg aggcggaaga tacctttggt ctgtatgaag atctggccca cagccagcgt
     2281 ggcgtaataa taaaactgga actgccgagt ggggccggat taaccgccga ctcgacgccg
     2341 ttaatgtatc aggggctgga agtcggacag ctgactaaac tggatttaaa tcctggtggt
     2401 aaagtcacag gggaaatgac cgttgatccc agcgtcgtta ccctgcttcg tgaaaatacc
     2461 cgcatcgaat tacgcaaccc gaaattatcc cttagcgatg ctaatctcag cgccctgctg
     2521 accggcaaaa ccttcgagct ggtgcccggc gatggcgagc cacgcaaaga attcgttgtt
     2581 gtgccaggcg aaaaagcact gctgcatgaa cctgatgttc tgacgctgac cctgaccgcg
     2641 ccggaaagtt acggtattga tgcgggtcag ccgctcattc ttcacggcgt gcaggtaggc
     2701 caggttatcg atcgtaaact caccagcaaa ggcgtcacct ttaccgtcgc catcgagcct
     2761 cagcatcgcg aactggtaaa aggcgatagc aaatttgtcg tcaacagccg tgtcgatgtg
     2821 aaggtggggc tggatggcgt tgagtttctc ggtgccagcg cctcagaatg gatcaatggc
     2881 gggatccgta ttctgccggg cgataaaggt gagatgaaag ccagctatcc actgtatgcc
     2941 aatttggaaa aagcgctgga gaacagcctt agcgatttac ccaccacaac cgtgagtttg
     3001 agtgcagaga cgctgccgga tgtgcaggca ggatcggtag tgctgtaccg taaatttgaa
     3061 gttggtgaag tgattaccgt gcgtccgcga gctaacgcgt ttgacatcga tctgcatatt
     3121 aagccggagt atcgcaacct tctgaccagc aatagcgtgt tctgggcaga aggcggggcg
     3181 aaagttcagc tgaatggtag tggtctgacc gtacaggcat ccccgctctc cagagctcta
     3241 aagggagcca ttagcttcga caacctcagc ggtgccagcg ccagtcagcg taaaggcgat
     3301 aagcgtattc tgtatgcttc cgaaacagcg gcccgtgcgg ttggcgggca gattacgctt
     3361 cacgctttcg atgccggaaa actggcggtc gggatgccaa ttcgctatct cggtattgac
     3421 atcgggcaaa tccagacgct ggatctgatt accgcacgca atgaagtaca ggcaaaggcg
     3481 gtactttacc cggaatatgt ccagaccttt gctcgcggtg gtacgcgctt ctcagtggtc
     3541 acaccgcaaa tttcggcagc tggcgttgag catcttgata ctatcctcca gccgtatatc
     3601 aacgtcgaac caggccgggg caatcctcgc cgcgactttg aattacaaga ggccaccatt
     3661 actgattcgc ggtacctgga tggcttaagc attattgttg aagcgccgga agccggttcg
     3721 ttaggcatcg gtacgcctgt gctgttccgt ggtctggaag tcggtacggt tacaggaatg
     3781 acgctgggga cattgtcaga tcgcgtgatg attgcgatgc gcatcagtaa acgctatcaa
     3841 cacctggtgc gtaacaattc cgtcttctgg ttggcatcgg gttacagtct ggactttggt
     3901 ctgacgggcg gcgtagtgaa aaccggcacc tttaaccagt ttatccgtgg cggcatcgcc
     3961 ttcgccacgc ctccgggtac gccactggca ccgaaagccc aggaaggcaa acacttcctg
     4021 ttgcaggaaa gtgaaccgaa agagtggcgt gaatggggaa ctgcgcttcc caaataatgc
     4081 ccacctcgag cagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     4141 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     4201 ttgctgaaag gaggaactat atccggataa ttcttgaaga cgaaagggcc tcgtgatacg
     4261 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     4321 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4381 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4441 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4501 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4561 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4621 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4681 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4741 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4801 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4861 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4921 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4981 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5041 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5101 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5161 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5221 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5281 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     5341 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5401 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5461 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5521 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5581 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     5641 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5701 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5761 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5821 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5881 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5941 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6001 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6061 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6121 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6181 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     6241 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc
     6301 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     6361 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     6421 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     6481 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     6541 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     6601 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     6661 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
     6721 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
     6781 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
     6841 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
     6901 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
     6961 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
     7021 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgc
//
