LOCUS       sequence-421618-        5810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Replicative vector for protein expression in Bacteroidales with an 
            aTc inducible promoter, terminator, and a barcoding site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5810)
  TITLE     Barcoded overexpression screens in gut Bacteroidales identify genes 
            with new roles in carbon utilization and stress resistance
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5810)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pNBU2_repA2
FEATURES             Location/Qualifiers
     source          1..5810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1349..1417,1418..2209)
                     /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     complement(1567..1586)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            2340..2449
                     /note="incP origin of transfer"
     rep_origin      2620..3008
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(3035..3658)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     3566..3585
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    3929..3947
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3991..4009
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 cgatgagaaa agactaatat ttatctatcc attcagtttg atttctcagg actttacatc
       61 gtcctgaaag tatttgttgg taccgaggac gcgtaaacat ttacagttgc atgtggccta
      121 ttgtttttag ccgttaaata ttttataact attaaatagc gatacaaatt gttcgaaact
      181 aatattgttt atatcatata ttctcgcatg ttttaaagct ttattaaatt gattttttgt
      241 aaacagtttt tcgtactctt tgttaaccca tttcattaca aaagtttcat atttttttct
      301 ctctttaaat gccatttttg ctggctttct ttttaataca attaatgtgc tatccacttt
      361 aggttttgga tggaaataat acctaggaat ttttgctaat atagaaatat ctacctctgc
      421 cattaacagc aatgctagtg atctgtttgt atctaataac attttagcaa aaccatattc
      481 cactattaaa taacttattg tggctgaact ttcaaaaaca atttttcgaa ttatatttgt
      541 gcttatgttg taaggtatgc tgccaaatat tttatatgga ttgtggctag gaaatgtaaa
      601 tttcagtata tcatcattta ctatttgata gttaggataa tttaagagct tattacgagt
      661 tacctcacat aatttagaat caatttctat cgccgttaca aaattacatc tctttaccaa
      721 tccagcagta aaatgacctt tccctgcacc tatttcaaag atgttatctt tttcatctaa
      781 acttatgcaa ttcattattt tttctatgtg atattttgaa gtaataaaat tttgactatc
      841 ttttatattt actttgttca ttataacctc tccttaattt attgcatctc ttttcgaata
      901 tttatgtttt ttgagaaaag aacgtactca tggttcatcc cgatatgcgt atcggtctgt
      961 atatcagcaa ctttctatgt gtttcaacta caatagtcat ctattctcat ctttctgagt
     1021 ccaccccctg caaagcccct ctttacgaca taaaaattcg gtcggaaaag gtatgcaaaa
     1081 gatgtttctc tctttaagag aaactcttcg ggatgcaaaa atatgaaaat aactccaatt
     1141 caccaaatta tatagcgact tttttacaaa atgctaaaat ttgttgattt ccgtcaagca
     1201 attgttgagc aaaaatgtct tttacgataa aatgatacct caatatcaac tgtttagcaa
     1261 aacgatattt ctcttaaaga gagaaacacc tttttgttca ccaatccccg acttttaatc
     1321 ccgcggccat gattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     1381 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     1441 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     1501 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     1561 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     1621 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1681 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1741 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     1801 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1861 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     1921 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1981 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     2041 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     2101 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     2161 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     2221 agtttactca taacgcgtca attcgagggg gatcaattcc gtgataggtg ggctgccctt
     2281 cctggttggc ttggtttcat cagccatccg cttgccctca tctgttacgc cggcggtagc
     2341 cggccagcct cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat aagggacagt
     2401 gaagaaggaa cacccgctcg cgggtgggcc tacttcacct atcctgcccg gctgacgccg
     2461 ttggatacac caaggaaagt ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa
     2521 aaaggatgga tataccgaaa aaatcgctat aatgaccccg aagcagggtt atgcagcgga
     2581 aaacggaatt gatccggcca cgatgcgtcc ggcgtagagg atctgaagat cagcagttca
     2641 acctgttgat agtacgtact aagctctcat gtttcacgta ctaagctctc atgtttaacg
     2701 tactaagctc tcatgtttaa cgaactaaac cctcatggct aacgtactaa gctctcatgg
     2761 ctaacgtact aagctctcat gtttcacgta ctaagctctc atgtttgaac aataaaatta
     2821 atataaatca gcaacttaaa tagcctctaa ggttttaagt tttataagaa aaaaaagaat
     2881 atataaggct tttaaagctt ttaaggttta acggttgtgg acaacaagcc agggatgtaa
     2941 cgcactgaga agcccttaga gcctctcaaa gcaattttga gtgacacagg aacacttaac
     3001 ggctgacatg ggaattcccc tccaccgcgg tggcttaaga cccactttca catttaagtt
     3061 gtttttctaa tccgcatatg atcaattcaa ggccgaataa gaaggctggc tctgcacctt
     3121 ggtgatcaaa taattcgata gcttgtcgta ataatggcgg catactatca gtagtaggtg
     3181 tttccctttc ttctttagcg acttgatgct cttgatcttc caatacgcaa cctaaagtaa
     3241 aatgccccac agcgctgagt gcatataatg cattctctag tgaaaaacct tgttggcata
     3301 aaaaggctaa ttgattttcg agagtttcat actgtttttc tgtaggccgt gtacctaaat
     3361 gtacttttgc tccatcgcga tgacttagta aagcacatct aaaactttta gcgttattac
     3421 gtaaaaaatc ttgccagctt tccccttcta aagggcaaaa gtgagtatgg tgcctatcta
     3481 acatctcaat ggctaaggcg tcgagcaaag cccgcttatt ttttacatgc caatacaatg
     3541 taggctgctc tacacctagc ttctgggcga gtttacgggt tgttaaacct tcgattccga
     3601 cctcattaag cagctctaat gcgctgttaa tcactttact tttatctaat ctagacatat
     3661 tcgtttaata tcataaataa tttattttat tttaaaatgc gcgggtgcaa aggtaagagg
     3721 ttttatttta actaccaaat gttttcggaa gttttttcgc ttttcttttt ctatcgtttc
     3781 tcagactctc ttagcgaaag ggaaagaagg taaagaagaa aaacaaaacg ccttttcttt
     3841 ttttgcaccc gctttccaag agaagaaagc cttgttaaat tgacttagtg taaaagcgca
     3901 gtactgcttg accataagaa caaaaaaatc tctatcactg atagggataa agtttggaag
     3961 ataaagctaa aagttcttat ctttgcagtc tccctatcag tgatagagag gaaataaaga
     4021 catataaaag aaaagacacc atgcacgtga gcctcggtac caaattccag aaaagaggcc
     4081 tcccgaaagg ggggcctttt ttcgttttgg tccgcctcgt gagagctgag agacgacatg
     4141 ttactatgct ccataacttc cgtctcttat cagttttttt gatgaagcga cctctataaa
     4201 ccaaaaaaga aacactccat tgctttacgg taatttttag tttcttttta gttactttta
     4261 cggcatagca taacacattg tgctgtaata tcttacaaag gtcatctacg acaaatttcc
     4321 gagcatctac gacaaatttc cgagcatcta cgacaaattt ccgaaaaaaa cggttatcta
     4381 cgacaaatat tctacgccat atatttgttt ttgtcgtaga taaatatttc ctttgcgaaa
     4441 agcattaaag tcatggagaa gaaagacaag aaacagaggg atatagtcct gtcgcaagac
     4501 aatgcgctga ctaccgcacg ctacaagttc gaccttatcg aaaaacgtgc cctatactcc
     4561 attatacgaa atatccgggc gcaatacatc gagaacgaga acggcagacg ggacttattc
     4621 gatgatctga ttatcaccat acacgaggaa gaactaaaga aatgcggtga caataccaag
     4681 atgcaaaaca tctacgatgc gctaaaacgg cttagaaatc gcgatataga gattaacaac
     4741 gaaaaaatgt ggctaagtat cggttttgtc aattacgtca agcatatcaa aaataaatcg
     4801 tattttgagg tgcaagtttc caaagaggta ctaccctatt acgtggaact cgcccaaaac
     4861 ttcacgagtt actccatgac ggtagcaatc gcactcaaaa gcacattctc gcaaaggttt
     4921 tacgaactgt gctgccaata ccgaaacgcc gggttctttt tctacacagt cgaaaagctt
     4981 cgggaaatgt tcatgttgga aaagaaatac aaacggggat gcgatttcaa aaggcgtgta
     5041 cttgacgatc cgcaaaagga actccgggag ttgtacaatg ccggacaatg cgatttatac
     5101 ttcgagttcg agccaaagag ctacaaaggc aaggaagtta ccgaatacaa attctacgtt
     5161 tacacaaggc aaaccgaaca acaaaagcta ttggaatacg aaagcgcaaa acaggcgatc
     5221 atctacataa caaccactat ttcgaggttc tctaaaagtg acaaaggata tgttaagcgt
     5281 gtagtgaacg ccgtacagtt acacccggaa atagccgtac aggtggcgca aaagatacaa
     5341 aagaagtttg agcaatatat agacaagcca gccaaacaga taggggctat catccgggtt
     5401 tgcttaatgg aagattttgg tatagaataa aaaacataca agtatgaaaa caggtgataa
     5461 agttatcgta ccagccgaaa taaacggcta tggcagagat ttacgggcaa tagttacaga
     5521 gatagagaag ttcgccgggg caatattcgt aacagtgata ttcaccgaac catgtccgga
     5581 agcctgcgga agaagggggg gggtgtttac catgacttcc agctaatagc aaggtgattt
     5641 attttgctat aatcgcaata aattgcaata atatctgtga taaattgcta taataatact
     5701 tataactacc tttgcaacag aatatagcat tttacagcaa tggggaaagg acaggaaagc
     5761 aagcaaaaga ggaaaacacc gcttaccacc atagcgatat taaaagaatc
//
