LOCUS       sequence-392030-        5842 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSN18 derivative encoding N. gonorrhoeae PglO with C-terminal FLAG 
            epitope tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5842)
  AUTHORS   Hershewe JM, Warfel KF, Iyer SM, Peruzzi JA, Sullivan CJ, Roth EW, 
            DeLisa MP, Kamat NP, Jewett MC
  TITLE     Improving cell-free glycoprotein synthesis by characterizing and 
            enriching native membrane vesicles.
  JOURNAL   Nat Commun. 2021 Apr 22;12(1):2363. doi: 10.1038/s41467-021-22329-3.
  PUBMED    33888690
REFERENCE   2  (bases 1 to 5842)
  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: "Nat 
            Commun."; date: "2021-04-22"; volume: "12(1):2363. doi"; pages: " 
            10.1038/s41467-021-22329-3"
COMMENT     Sequence Label: pSF-NgPglO
FEATURES             Location/Qualifiers
     source          1..5842
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1850..1873
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1938..1955)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2108..2194
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2286..2313
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2332..2423
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2424..2492,2493..3284)
                     /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(2642..2661)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3455..4043
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3944..3963
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4197..4214
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4229..4369
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4295..4314)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4455..4477
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4633..5511)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        5538..5822
                     /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     5745..5764
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 ggaggaatta accatggcat ctagtatgag cgcagaaacc accgttagcg gtgcacgtcc
       61 ggcagcaaaa ctgccgattt atatcctgcc gtgttttctg tggattggca ttattccgtt
      121 tacctttgca ctgcgtctga aaccgagtcc ggatttttat catgatgcag cagcagccgc
      181 aggtctgatt gttctgctgt ttctgaccgc aggtaaaaaa ctgttcgatg ttaaaattcc
      241 ggcaatcagc ttcctgctgt ttgcaatggc agcattttgg tggctgcagg cacgtctgat
      301 gaatctgatt tatccgggta tgaatgatat tgccagctgg gtttttattc tgctggcagt
      361 tagcgcatgg gcatgtaaaa gcctggttgc acattatggt caagaacgta ttgttaccct
      421 gtttgcatgg tcactgctga ttggtagcct gctgcagagc tgtattgttg ttattcagtt
      481 tgcaggctgg gaaaatacac cgctgctgca aaacattatt gttcatcgtg gtcagggtgt
      541 gattggtcat attggtcagc gtaataatct gggccattat ctgatgtggg gtattctggc
      601 aagcgcatat ctgaatggtc agcgcaaaat tcctgcagca ctgggtgcaa tttgtctgat
      661 tatgcagacc gcagttctgg gtctggttaa tagccgtacc attctgacct atattgcagc
      721 aattgcactg attctgccgt tttggtattt tcgtagcgat aaaagcaatc gtcgtaccat
      781 gctgggtatt gccgcagcag tttttctgac agcactgttt cagtttagca tgaatgcaat
      841 cctggaaacc tttaccggta ttcgttatga aaccgcagtt gaacgtgttg caaatggtgg
      901 ttttaccgat ctgcctcgtc agagcgaatg gaataaagca ctggcagcct ttcagagcgc
      961 accgattttt ggtcatggtt ggaatagctt tgcacagcag acctttctga ttaatgccga
     1021 acagcatacc atccacgata attttctgag caccctgttt acccatagcc ataacattat
     1081 tctgcagctg ctggccgaaa tgggtattag cggcaccctg ctggttgcag caaccctgct
     1141 gaccggtatt gcgggtctgc tgaaacgtag cctgacaccg gcaagcctgt ttctgctgtg
     1201 tgcactggcc gttagcatgt gtcatagcat gctggaatat ccgctgtggt atgtttattt
     1261 tctgattccg tttggcctga tgctgttcct gagccctgcc gaagcaagtg atggtattgc
     1321 cttcaaaaaa gcagcaaatc tgggtatcct gaccgcaagc gcagcaattt ttgcaggact
     1381 gctgcacctg gattggacct atacccgtct ggtgaatagt ttttcaccgg cagcagatga
     1441 tagcgcagaa accctgaatc gtaaaatcaa tgaactgcgc tatattagcg ccaatagccc
     1501 gatgctgagc ttttatgcag attttagcct ggttaatttt gccctgccgg aatatcctga
     1561 aacccagacc tgggcagaag aagccaccct gaaagcactg aaatatcgtc cgtatagcgc
     1621 aacctatcgt attgcactgt atctgatgcg tcagggtaaa gttgcggaag caaaacagtg
     1681 gatgcgtgca acccagagct attatccgta cctgatgcct cgttatgccg atgaaattcg
     1741 taaactgccg gtttgggcac ctctgctgcc agaactgctg aaagattgta aagcatttgc
     1801 agccgcaccg ggtcatccgg aaaccaaacc gtgtaaacct gcaggagcag actacaagga
     1861 cgatgacgac aagggataag tcgacgttta aacggtctcc agcttggctg ttttggcgga
     1921 tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa
     1981 cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa
     2041 gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc
     2101 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     2161 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc
     2221 gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat
     2281 taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat
     2341 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2401 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2461 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2521 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2581 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     2641 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     2701 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2761 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2821 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2881 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     2941 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     3001 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3061 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3121 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3181 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3241 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     3301 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3361 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3421 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3481 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3541 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3601 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3661 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3721 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3781 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3841 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3901 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3961 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4021 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4081 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4141 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     4201 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     4261 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     4321 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
     4381 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4441 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4501 cgcgaggcag cagatcaatt cgcgcgcgaa ggcgaagcgg catgcataat gtgcctgtca
     4561 aatggacgaa gcagggattc tgcaaaccct atgctactcc gtcaagccgt caattgtctg
     4621 attcgttacc aattatgaca acttgacggc tacatcattc actttttctt cacaaccggc
     4681 acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg
     4741 atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag
     4801 cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg
     4861 ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc
     4921 gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg
     4981 attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg
     5041 ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat
     5101 gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc
     5161 cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa
     5221 gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc
     5281 tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc cctgattttt
     5341 caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg
     5401 gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg
     5461 ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca tacttttcat
     5521 actcccgcca ttcagagaag aaaccaattg tccatattgc atcagacatt gccgtcactg
     5581 cgtcttttac tggctcttct cgctaaccaa accggtaacc ccgcttatta aaagcattct
     5641 gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg
     5701 cagaaaagtc cacattgatt atttgcacgg cgtcacactt tgctatgcca tagcattttt
     5761 atccataaga ttagcggatc ctacctgacg ctttttatcg caactctcta ctgtttctcc
     5821 atacccgttt tttgggctaa ca
//
