LOCUS       sequence-476620-        6031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FabG [3-oxoacyl-[acyl-carrier-protein] reductase] from 
            Yersinia pestis with His tag in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6031)
  TITLE     Josh Beckham Lab plasmids 2025
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6031)
  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: 
            "Unpublished"
COMMENT     Sequence Label: YpFabG_pET-28a(+)-TEV
FEATURES             Location/Qualifiers
     source          1..6031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     858..877
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(897..974)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1180..1199
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1283..1302
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1283..1301
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1302..1326
                     /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             1341..1363
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1382..1399
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1409..1429
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2174..2191
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2244..2262)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2258..2305
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2342..2797
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2429..2448)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2639..2660
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2890..3705)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2890..3705)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3613..3632
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3697..3716
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3827..4415
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4316..4335
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4569..4586
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4601..4743
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4669..4688)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4829..4851
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4845..5036)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5845..6031,1..896))
                     /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"
ORIGIN
        1 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
       61 ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
      121 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
      181 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
      241 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
      301 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
      361 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
      421 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
      481 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
      541 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
      601 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
      661 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
      721 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
      781 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
      841 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
      901 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
      961 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca
     1021 gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga
     1081 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     1141 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     1201 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
     1261 cgagatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac
     1321 aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacca tgggcagcag
     1381 ccatcatcat catcatcaca gcagcggcga gaatctttat tttcagggcc atatgtcttt
     1441 cgagggtaag atcgcactgg tcacaggggc atcgcgtgga attggacgcg ctatcgctga
     1501 gctgcttgtc gaacgtggag cgtgcgttat cgggacggct acctcggaaa agggtgctga
     1561 agctatcagt gcgtaccttg gcgaaaacgg gaaaggcctg atgttaaacg tagtcgatcc
     1621 cacatcaatc gacactgtac tggcaacaat tcgtgccgag ttcggagaag ttgacatcct
     1681 tgttaataat gcaggaatca cccgtgacaa cttgcttatg cgtatgaagg atgacgaatg
     1741 gcaagacatt attgatacaa acttgacttc ggtctttcgt ctgtcaaagg cagtgatgcg
     1801 tgcaatgatg aagaagcgtt tcggacgtat tatcactatt ggaagcgtag ttgggacgat
     1861 gggtaatgcg ggtcaggtta attatgcagc agctaaagcc ggagttattg gattctcgaa
     1921 aagcttggca cgcgaggtgg cttcgcgcgg aatcacagtc aacgttgtgg cgccaggctt
     1981 tatcgagacc gatatgacgc gcactttaac tgatgaccaa cgtgcaggaa tcttggcaca
     2041 ggtccccgcg aaccgcctgg gagacgcgaa agaaattgct tctgcagttg catttctggc
     2101 ctctgatgag gcatcgtaca tttctggaga gacgttacac gtcaatgggg gtatgtatat
     2161 gatctagctc gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa
     2221 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     2281 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg
     2341 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2401 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2461 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2521 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2581 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2641 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2701 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2761 aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt tcggggaaat
     2821 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2881 aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     2941 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     3001 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     3061 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     3121 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
     3181 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     3241 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     3301 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     3361 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     3421 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     3481 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     3541 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     3601 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     3661 taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     3721 actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgagtttt
     3781 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3841 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3901 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3961 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4021 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4081 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4141 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4201 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4261 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4321 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4381 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4441 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4501 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4561 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     4621 ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta caatctgctc
     4681 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     4741 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4801 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4861 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
     4921 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
     4981 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat
     5041 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg
     5101 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
     5161 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
     5221 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
     5281 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg
     5341 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
     5401 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     5461 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg
     5521 ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg
     5581 agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga
     5641 aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg
     5701 ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag
     5761 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     5821 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5881 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     5941 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     6001 ctgagagagt tgcagcaagc ggtccacgct g
//
