LOCUS       sequence-496778-        7576 bp    DNA     circular SYN 09-JAN-2026
DEFINITION  pLacI25 driving sfGFP in pNBU2 integrative Bacteroides plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7576)
  TITLE     Tropini Lab plasmids
REFERENCE   2  (bases 1 to 7576)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 9, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pEM404
FEATURES             Location/Qualifiers
     source          1..7576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(2..44)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             260..1339
                     /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
                     ALADSLMQLARQVSRLEET"
     primer_bind     complement(279..298)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      1440..1526
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    1639..1655
                     /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)."
     CDS             1793..2506
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             2513..2584
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             2588..2605
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2628..2655
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(join(4616..5407,5408..5476))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5239..5258
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6768..7156
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            7327..7436
                     /note="incP origin of transfer"
ORIGIN
        1 gataaaacga aaggctcagt cgaaagactg ggcctttcgt tttacaattg ggctaccttt
       61 tttttgtttt gtttgcaatg gttaatctat tgttaaaatt taaagtttca cttgaacttt
      121 caaataatgt tcttatattt ccagtgtcga aagaaacaaa gtaggactga tcacacaaca
      181 tttaaaaaat aacattatga aagcactcga aaagagattc aaaggtaata ttaacaataa
      241 tttattttca atggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg
      301 tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac
      361 gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca
      421 acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca
      481 cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt
      541 ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct
      601 tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat
      661 tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac
      721 acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct
      781 ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc
      841 gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc
      901 ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa
      961 tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat
     1021 gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga
     1081 cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg
     1141 cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa
     1201 gggcaatcag ctgttgccag tttcactggt gaaaagaaaa accaccctgg cgcccaatac
     1261 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     1321 ccgactggaa gagacgtaag tggagtgacc ttaaatcaag gacgatatta atcggaagga
     1381 gtattcaacg tgatgaagtc gcagggttga cgtgggacgt ctctgatcgg atctccaggc
     1441 atataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     1501 tgaacgctct cctgagtagg acaaatccca aagcactgct cgctttgtta agacggccca
     1561 aagcactgct cgctttgtta agacggcaat tgggctacct tttttttgtt ttgtttgcaa
     1621 tggttaatct attgttaatt gtgagcggat aacaagaact ttcaaataat gttcttatat
     1681 ttgcagtgtc gaaagaaaca aagtaggact gatcacacaa catttaaaaa ataacattat
     1741 gaaagcactc gaaaagagat tcaaaggtaa tattaacaat aatttatttt caatgcgtaa
     1801 aggcgaagag ctgttcactg gtgtcgtccc tattctggtg gaactggatg gtgatgtcaa
     1861 cggtcataag ttttccgtgc gtggcgaggg tgaaggtgac gcaactaatg gtaaactgac
     1921 gctgaagttc atctgtacta ctggtaaact gccggttcct tggccgactc tggtaacgac
     1981 gctgacttat ggtgttcagt gctttgctcg ttatccggac catatgaagc agcatgactt
     2041 cttcaagtcc gccatgccgg aaggctatgt gcaggaacgc acgatttcct ttaaggatga
     2101 cggcacgtac aaaacgcgtg cggaagtgaa atttgaaggc gataccctgg taaaccgcat
     2161 tgagctgaaa ggcattgact ttaaagaaga cggcaatatc ctgggccata agctggaata
     2221 caattttaac agccacaatg tttacatcac cgccgataaa caaaaaaatg gcattaaagc
     2281 gaattttaaa attcgccaca acgtggagga tggcagcgtg cagctggctg atcactacca
     2341 gcaaaacact ccaatcggtg atggtcctgt tctgctgcca gacaatcact atctgagcac
     2401 gcaaagcgtt ctgtctaaag atccgaacga gaaacgcgat catatggttc tgctggagtt
     2461 cgtaaccgca gcgggcatca cgcatggtat ggatgaactg tacaaaggat ctgattacaa
     2521 ggacgatgat gataaggact acaaagatga tgatgacaaa gattataagg acgatgatga
     2581 taaaggtcat catcatcatc atcactaaat cctagctgat tagctcgaga aggccatcct
     2641 gacggatggc cttttttttg tatcgagacg actagtgtaa ttgcctatct tccagtgatg
     2701 gaacagcatt tgtgcattgg ctgcaacaat cagccttact tgtgcctgtt ctatttccga
     2761 accgaccgct tgtatgaatg catcaaaatt cgttttctct acgttggatt ccttgttgct
     2821 catattgtga tgataatttc tacaaatata gtcattggta actatctatg aaactgtttg
     2881 atacttttat agttgattaa acttgttcat ggcatttgcc ttaatatcat ccgctatgtc
     2941 aatgtagggt ttcatagctt tgtagtcgct gtgtcccgtc catttcatga ccacctgtgc
     3001 cgggattccg agagccagcg cattgcagat gaatgtcctt tttcctgcat gggtactgag
     3061 caaagcgtat ttgggtgtga cttcatcaat acgttcattt cccttgtagt aggtttcccg
     3121 tacaggctcg ttgatttctg ccagttcgcc cagctctttc aggtaatcgt tcatcttctg
     3181 gttgctgatg acgggcagag ccatgtaatt ctcgaaatgg atgtccttgt atttgtccag
     3241 tatggctttg ctgtatttgt tcagttcaat cgtcaggctg tcggcagtct tgactgtggt
     3301 tatttcgatg tggtcggact tcacatcgct tcttttcaga ttgcgaacat ccgaataccg
     3361 caaactcgta aagcagcaga acaggaaaac atcacgcaca cgttccaggt attgcttatc
     3421 cttgggtatc tggtagtctt tcagcttgtt cagttcatcc caagtcagga agattacttt
     3481 tttcgaggtg gttttcagtt tcggtttgaa cgtatcgtat gcaatgttct gatgatgtcc
     3541 tttcttgaag ctccagcgca ggaaccattt gaggaatccc atttgcttgc cgatggtgct
     3601 gtttctcata tccttggtgt cacgcaggaa gttgacgtat tcgttcaatc caaactcgtt
     3661 gaaatagttg aacgttgcat cctccttgaa ctctttgagg tggttcctca ctgctgcaaa
     3721 tttttcatag gtggatgccg tccagttatt ctggttaccg cactctttta caaactcatc
     3781 gaacacctcc caaaagctga caggggcttc ttccggctgt tcttcactgg tgtctttcat
     3841 tctcatgttg aaagcttcct tcaactgttg ggtcgttggc atgacctcct gcacctcaaa
     3901 ttccttgaaa atattctgga tttcggcata gtatttcagc aagtccgtat tgatttcggc
     3961 tgcactttgc tttagcttgt tggtacatcc gttctttacc cgctgcttat ctgcatccca
     4021 tttggctacg tcaatccggt agcccgttgt aaactcgata cgttggctgg caaagatgac
     4081 acgcatacgg atgggtacgt tctctacgat tggcacaccg ttctttttcc ggctctccaa
     4141 tgcaaaaatg atgttgcgct tgatattcat aattgggtgc gtttgaaatt ctacacccaa
     4201 atatacaccc aattattgag atagcaaaag acatttagaa acatttactt ttactctata
     4261 ttgtaattta cacttgatta tcagtcgttt gcagttttat gatattctgt gaaagtataa
     4321 gttcgagagc ctgtctctcc gcaaaaaacg ctgaaaatca gcagattgca aaacaaacac
     4381 cctgttttac acccaagaat gtaaagtcgg gtgtttttgt tttatttaag ataatacaac
     4441 cactacataa taaaagagta gcgatattaa aagaatccga tgagaaaaga ctaatattta
     4501 tctatccatt cagtttgatt tctcaggact ttacatcgtc ctgaaagtat ttgttggtac
     4561 cggtaccgag gacgcgacag ctgcagacgc gttatgagta aacttggtct gacagttacc
     4621 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4681 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4741 ctgcaatgat accgcgggac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4801 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4861 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4921 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4981 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5041 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5101 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5161 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5221 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5281 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5341 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5401 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5461 aatgttgaat actcatactc ttcctttttc aatcatggcc gcgggattaa aagtcgggga
     5521 ttggtgaaca aaaaggtgtt tctctcttta agagaaatat cgttttgcta aacagttgat
     5581 attgaggtat cattttatcg taaaagacat ttttgctcaa caattgcttg acggaaatca
     5641 acaaatttta gcattttgta aaaaagtcgc tatataattt ggtgaattgg agttattttc
     5701 atatttttgc atcccgaaga gtttctctta aagagagaaa catcttttgc ataccttttc
     5761 cgaccgaatt tttatgtcgt aaagaggggc tttgcagggg gtggactcag aaagatgaga
     5821 atagatgact attgtagttg aaacacatag aaagttgctg atatacagac cgatacgcat
     5881 atcgggatga accatgagta cgttcttttc tcaaaaaaca taaatattcg aaaagagatg
     5941 caataaatta aggagaggtt ataatgaaca aagtaaatat aaaagatagt caaaatttta
     6001 ttacttcaaa atatcacata gaaaaaataa tgaattgcat aagtttagat gaaaaagata
     6061 acatctttga aataggtgca gggaaaggtc attttactgc tggattggta aagagatgta
     6121 attttgtaac ggcgatagaa attgattcta aattatgtga ggtaactcgt aataagctct
     6181 taaattatcc taactatcaa atagtaaatg atgatatact gaaatttaca tttcctagcc
     6241 acaatccata taaaatattt ggcagcatac cttacaacat aagcacaaat ataattcgaa
     6301 aaattgtttt tgaaagttca gccacaataa gttatttaat agtggaatat ggttttgcta
     6361 aaatgttatt agatacaaac agatcactag cattgctgtt aatggcagag gtagatattt
     6421 ctatattagc aaaaattcct aggtattatt tccatccaaa acctaaagtg gatagcacat
     6481 taattgtatt aaaaagaaag ccagcaaaaa tggcatttaa agagagaaaa aaatatgaaa
     6541 cttttgtaat gaaatgggtt aacaaagagt acgaaaaact gtttacaaaa aatcaattta
     6601 ataaagcttt aaaacatgcg agaatatatg atataaacaa tattagtttc gaacaatttg
     6661 tatcgctatt taatagttat aaaatattta acggctaaaa acaataggcc acatgcaact
     6721 gtaaatgttt acgcgggtac cgacaccgcg gtggagggga attcccatgt cagccgttaa
     6781 gtgttcctgt gtcactcaaa attgctttga gaggctctaa gggcttctca gtgcgttaca
     6841 tccctggctt gttgtccaca accgttaaac cttaaaagct ttaaaagcct tatatattct
     6901 tttttttctt ataaaactta aaaccttaga ggctatttaa gttgctgatt tatattaatt
     6961 ttattgttca aacatgagag cttagtacgt gaaacatgag agcttagtac gttagccatg
     7021 agagcttagt acgttagcca tgagggttta gttcgttaaa catgagagct tagtacgtta
     7081 aacatgagag cttagtacgt gaaacatgag agcttagtac gtactatcaa caggttgaac
     7141 tgctgatctt cagatcctct acgccggacg catcgtggcc ggatcaattc cgttttccgc
     7201 tgcataaccc tgcttcgggg tcattatagc gattttttcg gtatatccat cctttttcgc
     7261 acgatataca ggattttgcc aaagggttcg tgtagacttt ccttggtgta tccaacggcg
     7321 tcagccgggc aggataggtg aagtaggccc acccgcgagc gggtgttcct tcttcactgt
     7381 cccttattcg cacctggcgg tgctcaacgg gaatcctgct ctgcgaggct ggccggctac
     7441 cgccggcgta acagatgagg gcaagcggat ggctgatgaa accaagccaa ccaggaaggg
     7501 cagcccacct atcacggaat tgatccccct cgaattggaa ttcgttcctc gattcagatc
     7561 taaaggatcc aatcta
//
