LOCUS       sequence-334333-        4390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TESEC expression vector for mEGFP-Alr fusion construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4390)
  AUTHORS   Bongaerts N, Edoo Z, Abukar AA, Song X, Sosa-Carrillo S, 
            Haggenmueller S, Savigny J, Gontier S, Lindner AB, Wintermute EH
  TITLE     Low-cost anti-mycobacterial drug discovery using engineered E. coli.
  JOURNAL   Nat Commun. 2022 Jul 7;13(1):3905. doi: 10.1038/s41467-022-31570-3.
  PUBMED    35798732
REFERENCE   2  (bases 1 to 4390)
  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: "2022-07-7"; volume: "13(1):3905. doi"; pages: " 
            10.1038/s41467-022-31570-3"
COMMENT     Sequence Label: pNB203
FEATURES             Location/Qualifiers
     source          1..4390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      35..121
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(286..874)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(366..385)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(962..1056)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(1086..1877,1878..1946))
                     /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     1709..1728
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1947..2051)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2095..2379
                     /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     2302..2321
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             2390..2401
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(2405..2407,2408..2410,2411..3121)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2450..2471)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2711..2730)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3058..3079
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aggtgggatc ccatggtacg cgtgctagag gcatcaaata aaacgaaagg ctcagtcgaa
       61 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
      121 tccgccgccc tagacctagg gcgttcggct gcggcgagcg gtatcagctc actcaaaggc
      181 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
      241 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      301 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      361 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      421 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      481 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      541 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      601 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
      661 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
      721 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
      781 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
      841 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
      901 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga ctagtgcttg
      961 gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga
     1021 gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcgta aacttggtct
     1081 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     1141 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     1201 ggccccagtg ctgcaatgat accgcgggac ccacgctcac cggctccaga tttatcagca
     1261 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     1321 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     1381 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     1441 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     1501 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     1561 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     1621 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     1681 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     1741 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     1801 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     1861 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     1921 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     1981 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     2041 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgaagacat ggagaagaaa
     2101 ccaattgtcc atattgcatc agacattgcc gtcactgcgt cttttactgg ctcttctcgc
     2161 taaccaaacc ggtaaccccg cttattaaaa gcattctgta acaaagcggg accaaagcca
     2221 tgacaaaaac gcgtaacaaa agtgtctata atcacggcag aaaagtccac attgattatt
     2281 tgcacggcgt cacactttgc tatgccatag catttttatc cataagatta gcggatccta
     2341 cctgacgctt tttatcgcaa ctctctactg tttctccata gaattcatta aagaggagaa
     2401 aggaatggtt agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2461 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     2521 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2581 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     2641 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2701 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2761 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2821 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     2881 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     2941 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     3001 ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     3061 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     3121 gggtggtggt ggttctggtg gtggtggttc tggtggtggt ggttctaaac gcttttggga
     3181 aaatgtgggt aaacctaacg atacgacgga cggtcgcggc accacctccc tcgcgatgac
     3241 accgatctca cagactccgg gcttactggc cgaggcgatg gttgacctcg gagctattga
     3301 acataatgtg cgtgtcctcc gcgaacatgc gggccacgcc cagttgatgg cagtcgttaa
     3361 ggcagacggc tacggtcacg gggctacacg cgtagctcaa acggccttag gcgccggtgc
     3421 tgcggaactg ggggtcgcga cagtggatga agctcttgcg ctgcgtgccg acggcattac
     3481 cgcgccggta ttagcgtggc tgcatccgcc gggtatcgac tttggtccag ccctgctggc
     3541 cgatgttcag gtggcggttt cttcactgcg ccagttggat gaattgctgc acgcggttcg
     3601 ccgtaccggt cgcaccgcga ccgtcacggt caaagtggat accggactca accgcaacgg
     3661 cgtaggtcca gcacaattcc cagcgatgtt aacggcgctt cgtcaggcca tggctgaaga
     3721 tgctgtgcgc ctgcgcggcc tgatgagcca tatggtttat gcagataagc ccgacgatag
     3781 tattaatgac gtacaagcgc aacgttttac tgctttcctg gcgcaggcgc gtgaacaggg
     3841 cgtgcgtttt gaagtggcac atctgtcaaa cagttcggcc acgatggccc gcccagacct
     3901 gacattcgat ctggtgcgcc ctggtatcgc tgtctacggg ctttcaccgg tgccggccct
     3961 gggtgatatg ggcctggtgc cggcaatgac agtcaaatgt gccgtggcgc tcgtgaaaag
     4021 tattcgtgcg ggtgaaggcg tgtcttatgg gcatacatgg atcgcacccc gcgatactaa
     4081 tctggcgctg ttgccgattg gctatgccga tggcgttttt cgctcattag gcggacgtct
     4141 ggaagttctg attaatggac gccgctgccc aggtgtaggt cgtatttgta tggatcagtt
     4201 tatggtcgat ctgggtccgg gcccgctgga cgtcgcggag ggtgacgaag cgatcctgtt
     4261 tggtccgggc atccgcggag aaccgaccgc ccaagactgg gcggacctgg tggggactat
     4321 tcactacgaa gtggtaacct caccgcgcgg gcgtattacc cgtacgtacc gcgaagctga
     4381 aaaccgctaa
//
