LOCUS       sequence-261694-        6977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of C. jejuni PglB and F. tularensis LpxE in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6977)
  AUTHORS   Stark JC, Jaroentomeechai T, Moeller TD, Hershewe JM, Warfel KF, 
            Moricz BS, Martini AM, Dubner RS, Hsu KJ, Stevenson TC, Jones BD, 
            DeLisa MP, Jewett MC
  TITLE     On-demand biomanufacturing of protective conjugate vaccines.
  JOURNAL   Sci Adv. 2021 Feb 3;7(6). pii: 7/6/eabe9444. doi: 
            10.1126/sciadv.abe9444. Print 2021 Feb.
  PUBMED    33536221
REFERENCE   2  (bases 1 to 6977)
  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 Adv."; 
            date: "2021-02-3"; volume: "7(6). pii: 7/6/eabe9444. doi"; pages: " 
            10.1126/sciadv.abe9444. Print 2021 Feb"
COMMENT     Sequence Label: pSF-CjPglB-LpxE
FEATURES             Location/Qualifiers
     source          1..6977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(214..233)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(215..233)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(243..259)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(252..274)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(424..1302)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1329..1613
                     /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     1536..1555
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3810..3833
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3898..3915)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4068..4154
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4246..4273
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4292..4383
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4384..4452,4453..5244)
                     /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(4602..4621)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5415..6003
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5904..5923
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6157..6174
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(6248..6253,6254..6271,6272..6278)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6286..6302
                     /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)."
     primer_bind     6291..6313
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6310..6326
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6310..6326
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6345..6365
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6347..6365
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     RBS             6414..6436
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 ttatagcatt tttttatatt ttccctaagc taagatattt ttggtatttg ctgatagtgg
       61 tatttgctgg gagtagaatt atggttggtt cacattatcc tagtgatgta atttttggcg
      121 ttgcttttgg ttgttactgt acagcatata tctactattg gatgagaaat agagagatta
      181 tttagctcga gggggggccc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc
      241 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
      301 cgcaaagccc gccgaaaggc gggctttttt ttatgcataa tgtgcctgtc aaatggacga
      361 agcagggatt ctgcaaaccc tatgctactc cgtcaagccg tcaattgtct gattcgttac
      421 caattatgac aacttgacgg ctacatcatt cactttttct tcacaaccgg cacggaactc
      481 gctcgggctg gccccggtgc attttttaaa tacccgcgag aaatagagtt gatcgtcaaa
      541 accaacattg cgaccgacgg tggcgatagg catccgggtg gtgctcaaaa gcagcttcgc
      601 ctggctgata cgttggtcct cgcgccagct taagacgcta atccctaact gctggcggaa
      661 aagatgtgac agacgcgacg gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa
      721 attgctgtct gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc gattatccat
      781 cggtggatgg agcgactcgt taatcgcttc catgcgccgc agtaacaatt gctcaagcag
      841 atttatcgcc agcagctccg aatagcgccc ttccccttgc ccggcgttaa tgatttgccc
      901 aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaacc ccgtattggc
      961 aaatattgac ggccagttaa gccattcatg ccagtaggcg cgcggacgaa agtaaaccca
     1021 ctggtgatac cattcgcgag cctccggatg acgaccgtag tgatgaatct ctcctggcgg
     1081 gaacagcaaa atatcacccg gtcggcaaac aaattctcgt ccctgatttt tcaccacccc
     1141 ctgaccgcga atggtgagat tgagaatata acctttcatt cccagcggtc ggtcgataaa
     1201 aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc gccaccagat gggcattaaa
     1261 cgagtatccc ggcagcaggg gatcattttg cgcttcagcc atacttttca tactcccgcc
     1321 attcagagaa gaaaccaatt gtccatattg catcagacat tgccgtcact gcgtctttta
     1381 ctggctcttc tcgctaacca aaccggtaac cccgcttatt aaaagcattc tgtaacaaag
     1441 cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg gcagaaaagt
     1501 ccacattgat tatttgcacg gcgtcacact ttgctatgcc atagcatttt tatccataag
     1561 attagcggat cctacctgac gctttttatc gcaactctct actgtttctc catacccgtt
     1621 ttttgggcta acaggaggaa ttaaccatgg catctagtat gttgaaaaaa gagtatttaa
     1681 aaaaccctta tttagttttg tttgcgatga ttatattagc ttatgttttt agtgtatttt
     1741 gcaggtttta ttgggtttgg tgggcaagtg agtttaatga gtattttttc aataatcagt
     1801 taatgatcat ttcaaatgat ggctatgctt ttgctgaggg cgcaagagat atgatagcag
     1861 gttttcatca gcctaatgat ttgagttatt atggatcttc tttatccgcg cttacttatt
     1921 ggctttataa aatcacacct ttttcttttg aaagtatcat tttatatatg agtacttttt
     1981 tatcttcttt ggtggtgatt cctactattt tgctagctaa cgaatacaaa cgtcctttaa
     2041 tgggctttgt agctgctctt ttagcaagta tagcaaacag ttattataat cgcactatga
     2101 gtgggtatta tgatacggat atgctggtaa ttgttttgcc tatgtttatt ttatttttta
     2161 tggtaagaat gattttaaaa aaagactttt tttcattgat tgccttgccg ttatttatag
     2221 gaatttatct ttggtggtat ccttcaagtt atactttaaa tgtagcttta attggacttt
     2281 ttttaattta tacacttatt tttcatagaa aagaaaagat tttttatata gctgtgattt
     2341 tgtcttctct tactctttca aatatagcat ggttttatca aagtgccatt atagtaatac
     2401 tttttgcttt attcgcctta gagcaaaaac gcttaaattt tatgattata ggaattttag
     2461 gtagtgcaac tttgatattt ttgattttaa gtggtggggt tgatcctata ctttatcagc
     2521 ttaaatttta tatttttaga agtgatgaaa gtgcgaattt aacgcagggc tttatgtatt
     2581 ttaatgtcaa tcaaaccata caagaagttg aaaatgtaga tcttagcgaa tttatgcgaa
     2641 gaattagtgg tagtgaaatt gtttttttgt tttctttgtt tggttttgta tggcttttga
     2701 gaaaacataa aagtatgatt atggctttac ctatattggt gcttgggttt ttagccttaa
     2761 aaggggggct tagatttacc atttattctg tacctgtaat ggccttagga tttggttttt
     2821 tattgagcga gtttaaggct ataatggtta aaaaatatag ccaattaact tcaaatgttt
     2881 gtattgtttt tgcaactatt ttgactttag ctccagtatt tatccatatt tacaactata
     2941 aagcgccaac agttttttct caaaatgaag catcattatt aaatcaatta aaaaatatag
     3001 ccaatagaga agattatgtg gtaacttggt gggattatgg ttatcctgtg cgttattata
     3061 gcgatgtgaa aactttagta gatggtggaa agcatttagg taaggataat tttttccctt
     3121 cttttgcttt aagcaaagat gaacaagctg cagctaatat ggcaagactt agtgtagaat
     3181 atacagaaaa aagcttttat gctccgcaaa atgatatttt aaaaacagac attttgcaag
     3241 ccatgatgaa agattataat caaagcaatg tggatttgtt tctagcttca ttatcaaaac
     3301 ctgattttaa aatcgatacg ccaaaaactc gtgatattta tctttatatg cccgctagaa
     3361 tgtctttgat tttttctacg gtggctagtt tttcttttat taatttagat acaggagttt
     3421 tggataaacc ttttaccttt agcacagctt atccacttga tgttaaaaat ggagaaattt
     3481 atcttagcaa cggagtggtt ttaagcgatg attttagaag ttttaaaata ggtgataatg
     3541 tggtttctgt aaatagtatc gtagagatta attctattaa acaaggtgaa tacaaaatca
     3601 ctccaattga tgataaggct cagttttata ttttttattt aaaggatagt gctattcctt
     3661 acgcacaatt tattttaatg gataaaacca tgtttaatag tgcttatgtg caaatgtttt
     3721 ttttaggaaa ttatgataag aatttatttg acttggtgat taattctaga gatgctaagg
     3781 tttttaaact taaaattcct gcaggagcag actacaagga cgatgacgac aagggataag
     3841 tcgacgttta aacggtctcc agcttggctg ttttggcgga tgagagaaga ttttcagcct
     3901 gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag
     3961 tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga
     4021 tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa
     4081 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     4141 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
     4201 ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga
     4261 cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaaat
     4321 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4381 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4441 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4501 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4561 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4621 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4681 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4741 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4801 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4861 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4921 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4981 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5041 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5101 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5161 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5221 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5281 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5341 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5401 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5461 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5521 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     5581 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5641 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5701 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5761 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5821 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5881 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5941 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6001 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6061 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6121 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6181 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     6241 tgcaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     6301 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     6361 aaagggaaca aaagctggag ctccaccgcg gtggcggccg ctctagaaat aattttgttt
     6421 aactttaaga aggagatata ccatggtcaa acagacatta caaacaaact ttcaaggttt
     6481 taaagatatt tttaaaaaac caaaactaca caatcataaa ttgcctagat atctacagtt
     6541 gaaatatacg tttataccat tattaatttt ggtaattttt gcatactata acttagatac
     6601 cccagttgag aactatatca agcattctat gccaaatatt gttggtgtaa tttttggtaa
     6661 aataactgat gttggtaagg ccgagtatat tttgataatt tgcggtgtga tagtgttagc
     6721 gcgtttattt acagatagcc aaaaattatc tgctaatact agagctatgt ttgacaaggt
     6781 gtcggcatat gcgggtttta tcttagcaac tgtagctatt agtggtattt tgggacaaat
     6841 actcaagatg ataataggta gagcgcgtcc taagtttttc ttggaatatg gttcgcatta
     6901 tttccaacat tttcatgcac ctggatatga ttttgcaagt atgccgtcag ggcactcaat
     6961 cacagttgga gcaatgt
//
