LOCUS       sequence-496774-        6714 bp    DNA     circular SYN 09-JAN-2026
DEFINITION  Entry Vector for inserting Inducible promoters upstream of sfGFP in 
            pNBU2 integrative Bacteroides plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 16 to 50)
  TITLE     Tropini Lab plasmids
REFERENCE   2  (bases 1 to 6714)
  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: pEM124
FEATURES             Location/Qualifiers
     source          1..6714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..50
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     terminator      816..843
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             964..1677
                     /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             1684..1755
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             1759..1776
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1799..1826
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(join(3787..4578,4579..4647))
                     /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     4410..4429
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5939..6327
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            6498..6607
                     /note="incP origin of transfer"
ORIGIN
        1 gttctgagac cctatttgac ggctagctca gtcctaggta cagtgctagc gtacgtactt
       61 taactttaag aaggagatat acataatggt ttcgaaaggg gaagaagata acatggctat
      121 cattaaggag tttatgcgct tcaaggtgca catggagggc tctgtcaacg gccacgagtt
      181 cgaaatcgaa ggtgaaggtg aaggtcgtcc gtacgaaggt actcagaccg caaagttgaa
      241 agttacaaaa ggaggtccgt tgcctttcgc ttgggacatt ctgagcccgc aatttatgta
      301 cgggagcaaa gcctatgtta aacatccggc ggatattccc gattacctga aactgtcatt
      361 cccggagggg ttcaagtggg aacgcgtcat gaactttgaa gacggtggcg tggttacagt
      421 gactcaggat tcgtcccttc aagatggcga atttatctac aaagtcaaac tgcgcgggac
      481 aaattttccg agtgacggtc ctgttatgca gaagaagacc atggggtggg aagcaagctc
      541 cgaacgcatg taccccgagg acggtgcact caaaggagaa atcaagcagc gtctgaaact
      601 gaaagatggc gggcactacg atgcagaagt gaaaaccacc tataaggcga aaaagcctgt
      661 acaactgccg ggtgcctata atgtgaacat taaactggat ataacgagtc acaacgaaga
      721 ctatacaatc gtcgagcagt atgagcgtgc ggaaggacgt cattccactg gtggaatgga
      781 tgaactgtat aagtaaatcc tagctgatta gctcgagaag gccatcctga cggatggcct
      841 tttttttgta tcgagacgac tagtgtaatt gcctatcttc cagtgatgga acagcatttg
      901 tgcattggct gcaacaatca gccttacttg tgcctgttct atttccgaac cgaccgggtc
      961 tcaatgcgta aaggcgaaga gctgttcact ggtgtcgtcc ctattctggt ggaactggat
     1021 ggtgatgtca acggtcataa gttttccgtg cgtggcgagg gtgaaggtga cgcaactaat
     1081 ggtaaactga cgctgaagtt catctgtact actggtaaac tgccggttcc ttggccgact
     1141 ctggtaacga cgctgactta tggtgttcag tgctttgctc gttatccgga ccatatgaag
     1201 cagcatgact tcttcaagtc cgccatgccg gaaggctatg tgcaggaacg cacgatttcc
     1261 tttaaggatg acggcacgta caaaacgcgt gcggaagtga aatttgaagg cgataccctg
     1321 gtaaaccgca ttgagctgaa aggcattgac tttaaagaag acggcaatat cctgggccat
     1381 aagctggaat acaattttaa cagccacaat gtttacatca ccgccgataa acaaaaaaat
     1441 ggcattaaag cgaattttaa aattcgccac aacgtggagg atggcagcgt gcagctggct
     1501 gatcactacc agcaaaacac tccaatcggt gatggtcctg ttctgctgcc agacaatcac
     1561 tatctgagca cgcaaagcgt tctgtctaaa gatccgaacg agaaacgcga tcatatggtt
     1621 ctgctggagt tcgtaaccgc agcgggcatc acgcatggta tggatgaact gtacaaagga
     1681 tctgattaca aggacgatga tgataaggac tacaaagatg atgatgacaa agattataag
     1741 gacgatgatg ataaaggtca tcatcatcat catcactaaa tcctagctga ttagctcgag
     1801 aaggccatcc tgacggatgg cctttttttt gtatcgagac gactagtgta attgcctatc
     1861 ttccagtgat ggaacagcat ttgtgcattg gctgcaacaa tcagccttac ttgtgcctgt
     1921 tctatttccg aaccgaccgc ttgtatgaat gcatcaaaat tcgttttctc tacgttggat
     1981 tccttgttgc tcatattgtg atgataattt ctacaaatat agtcattggt aactatctat
     2041 gaaactgttt gatactttta tagttgatta aacttgttca tggcatttgc cttaatatca
     2101 tccgctatgt caatgtaggg tttcatagct ttgtagtcgc tgtgtcccgt ccatttcatg
     2161 accacctgtg ccgggattcc gagagccagc gcattgcaga tgaatgtcct ttttcctgca
     2221 tgggtactga gcaaagcgta tttgggtgtg acttcatcaa tacgttcatt tcccttgtag
     2281 taggtttccc gtacaggctc gttgatttct gccagttcgc ccagctcttt caggtaatcg
     2341 ttcatcttct ggttgctgat gacgggcaga gccatgtaat tctcgaaatg gatgtccttg
     2401 tatttgtcca gtatggcttt gctgtatttg ttcagttcaa tcgtcaggct gtcggcagtc
     2461 ttgactgtgg ttatttcgat gtggtcggac ttcacatcgc ttcttttcag attgcgaaca
     2521 tccgaatacc gcaaactcgt aaagcagcag aacaggaaaa catcacgcac acgttccagg
     2581 tattgcttat ccttgggtat ctggtagtct ttcagcttgt tcagttcatc ccaagtcagg
     2641 aagattactt ttttcgaggt ggttttcagt ttcggtttga acgtatcgta tgcaatgttc
     2701 tgatgatgtc ctttcttgaa gctccagcgc aggaaccatt tgaggaatcc catttgcttg
     2761 ccgatggtgc tgtttctcat atccttggtg tcacgcagga agttgacgta ttcgttcaat
     2821 ccaaactcgt tgaaatagtt gaacgttgca tcctccttga actctttgag gtggttcctc
     2881 actgctgcaa atttttcata ggtggatgcc gtccagttat tctggttacc gcactctttt
     2941 acaaactcat cgaacacctc ccaaaagctg acaggggctt cttccggctg ttcttcactg
     3001 gtgtctttca ttctcatgtt gaaagcttcc ttcaactgtt gggtcgttgg catgacctcc
     3061 tgcacctcaa attccttgaa aatattctgg atttcggcat agtatttcag caagtccgta
     3121 ttgatttcgg ctgcactttg ctttagcttg ttggtacatc cgttctttac ccgctgctta
     3181 tctgcatccc atttggctac gtcaatccgg tagcccgttg taaactcgat acgttggctg
     3241 gcaaagatga cacgcatacg gatgggtacg ttctctacga ttggcacacc gttctttttc
     3301 cggctctcca atgcaaaaat gatgttgcgc ttgatattca taattgggtg cgtttgaaat
     3361 tctacaccca aatatacacc caattattga gatagcaaaa gacatttaga aacatttact
     3421 tttactctat attgtaattt acacttgatt atcagtcgtt tgcagtttta tgatattctg
     3481 tgaaagtata agttcgagag cctgtctctc cgcaaaaaac gctgaaaatc agcagattgc
     3541 aaaacaaaca ccctgtttta cacccaagaa tgtaaagtcg ggtgtttttg ttttatttaa
     3601 gataatacaa ccactacata ataaaagagt agcgatatta aaagaatccg atgagaaaag
     3661 actaatattt atctatccat tcagtttgat ttctcaggac tttacatcgt cctgaaagta
     3721 tttgttggta ccggtaccga ggacgcgaca gctgcagacg cgttatgagt aaacttggtc
     3781 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3841 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3901 tggccccagt gctgcaatga taccgcggga cccacgctca ccggctccag atttatcagc
     3961 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4021 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4081 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4141 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4201 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4261 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4321 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4381 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4441 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4501 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4561 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4621 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatcatggc cgcgggatta
     4681 aaagtcgggg attggtgaac aaaaaggtgt ttctctcttt aagagaaata tcgttttgct
     4741 aaacagttga tattgaggta tcattttatc gtaaaagaca tttttgctca acaattgctt
     4801 gacggaaatc aacaaatttt agcattttgt aaaaaagtcg ctatataatt tggtgaattg
     4861 gagttatttt catatttttg catcccgaag agtttctctt aaagagagaa acatcttttg
     4921 catacctttt ccgaccgaat ttttatgtcg taaagagggg ctttgcaggg ggtggactca
     4981 gaaagatgag aatagatgac tattgtagtt gaaacacata gaaagttgct gatatacaga
     5041 ccgatacgca tatcgggatg aaccatgagt acgttctttt ctcaaaaaac ataaatattc
     5101 gaaaagagat gcaataaatt aaggagaggt tataatgaac aaagtaaata taaaagatag
     5161 tcaaaatttt attacttcaa aatatcacat agaaaaaata atgaattgca taagtttaga
     5221 tgaaaaagat aacatctttg aaataggtgc agggaaaggt cattttactg ctggattggt
     5281 aaagagatgt aattttgtaa cggcgataga aattgattct aaattatgtg aggtaactcg
     5341 taataagctc ttaaattatc ctaactatca aatagtaaat gatgatatac tgaaatttac
     5401 atttcctagc cacaatccat ataaaatatt tggcagcata ccttacaaca taagcacaaa
     5461 tataattcga aaaattgttt ttgaaagttc agccacaata agttatttaa tagtggaata
     5521 tggttttgct aaaatgttat tagatacaaa cagatcacta gcattgctgt taatggcaga
     5581 ggtagatatt tctatattag caaaaattcc taggtattat ttccatccaa aacctaaagt
     5641 ggatagcaca ttaattgtat taaaaagaaa gccagcaaaa atggcattta aagagagaaa
     5701 aaaatatgaa acttttgtaa tgaaatgggt taacaaagag tacgaaaaac tgtttacaaa
     5761 aaatcaattt aataaagctt taaaacatgc gagaatatat gatataaaca atattagttt
     5821 cgaacaattt gtatcgctat ttaatagtta taaaatattt aacggctaaa aacaataggc
     5881 cacatgcaac tgtaaatgtt tacgcgggta ccgacaccgc ggtggagggg aattcccatg
     5941 tcagccgtta agtgttcctg tgtcactcaa aattgctttg agaggctcta agggcttctc
     6001 agtgcgttac atccctggct tgttgtccac aaccgttaaa ccttaaaagc tttaaaagcc
     6061 ttatatattc ttttttttct tataaaactt aaaaccttag aggctattta agttgctgat
     6121 ttatattaat tttattgttc aaacatgaga gcttagtacg tgaaacatga gagcttagta
     6181 cgttagccat gagagcttag tacgttagcc atgagggttt agttcgttaa acatgagagc
     6241 ttagtacgtt aaacatgaga gcttagtacg tgaaacatga gagcttagta cgtactatca
     6301 acaggttgaa ctgctgatct tcagatcctc tacgccggac gcatcgtggc cggatcaatt
     6361 ccgttttccg ctgcataacc ctgcttcggg gtcattatag cgattttttc ggtatatcca
     6421 tcctttttcg cacgatatac aggattttgc caaagggttc gtgtagactt tccttggtgt
     6481 atccaacggc gtcagccggg caggataggt gaagtaggcc cacccgcgag cgggtgttcc
     6541 ttcttcactg tcccttattc gcacctggcg gtgctcaacg ggaatcctgc tctgcgaggc
     6601 tggccggcta ccgccggcgt aacagatgag ggcaagcgga tggctgatga aaccaagcca
     6661 accaggaagg gcagcccacc tatcacggaa ttgatccccc tcgaattgga attc
//
