LOCUS       sequence-430541-        7573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NBU2 backbone with LacI(YQR) and P(O1)_sgRNA targeting Nanoluc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7573)
  AUTHORS   Huang BD, Groseclose TM, Wilson CJ
  TITLE     Transcriptional programming in a Bacteroides consortium.
  JOURNAL   Nat Commun. 2022 Jul 6;13(1):3901. doi: 10.1038/s41467-022-31614-8.
  PUBMED    35794179
REFERENCE   2  (bases 1 to 7573)
  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: "2022-07-6"; volume: "13(1):3901. doi"; pages: " 
            10.1038/s41467-022-31614-8"
COMMENT     Sequence Label: pBH502
FEATURES             Location/Qualifiers
     source          1..7573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    182..198
                     /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)."
     protein_bind    215..231
                     /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)."
     ncRNA           244..280
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     misc_RNA        301..376
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           383..450
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     CDS             874..1956
                     /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
                     EACKTAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(893..912)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             join(6017..6085,6086..6877)
                     /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(6235..6254)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            7008..7117
                     /note="incP origin of transfer"
     rep_origin      join(7288..7573,1..103)
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 gtttaacggt tgtggacaac aagccaggga tgtaacgcac tgagaagccc ttagagcctc
       61 tcaaagcaat tttgagtgac acaggaacac ttaacggctg acatgggaat tcccctccac
      121 cgcggtggcc ctgtacaaag aaaattcgac aaactgttat ttttctatct atttatttga
      181 attgtgagcg gataacaatt acctttgtcg gcaattgtga gcggataaca attatgatct
      241 gtcctgatga gtccgtgagg acgaaacgag taagctcgtc gacagaacga tgcgctgaat
      301 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
      361 ggcaccgagt cggtgctttt ttggccggca tggtcccagc ctcctcgctg gcgccggctg
      421 ggcaacatgc ttcggcatgg cgaatgggac gcactctaat cgttatcgga gtgcttttag
      481 attactaatc aaattgcttc tactaattgc ctatcttcca gtgatggaac agcatttgaa
      541 cgcattggct gcaacaatca gccttgatct ggaagaagca atgaaagctg ctgttaagtc
      601 tccgaatcag gtattgttcc tgacaggtgt attcccatcc ggtaaacgcg gatactttgc
      661 agttgatctg actcaggaat aaattataaa ttaaggtaag aagattgtag gataagctaa
      721 tgaaatagaa aaaggatgcc gtcacacaac ttgtcggcat tcttttttgt tttattagtt
      781 gaaaatatag tgaaaaagtt gcctaaatat gtatgttaac aaattatttg tcgtaacttt
      841 gcactccaaa tctgttttta acatatggca ctagtgaaac cagtaacgtt atacgatgtc
      901 gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac
      961 gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc
     1021 aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc
     1081 agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa
     1141 ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaaacg
     1201 gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat
     1261 gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat
     1321 gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg
     1381 ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta
     1441 agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa
     1501 attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc
     1561 atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg
     1621 gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg
     1681 gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc
     1741 aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag
     1801 ggccaggcgg tgaagggcaa tcagctgttg cccgtatcac tggtgaaaag aaaaaccacc
     1861 ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     1921 gcacgacagg tttcccgact ggaaagcggg cagtgagctt tcctcggtac caaattccag
     1981 aaaagaggcc tcccgaaagg ggggcctttt ttcgttttgg tcctacttgt gcctgttcta
     2041 tttcctatgc ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2101 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac atcccaaatg
     2161 ctgttccatc actggaagat aggcaattag tagaagcaat ttgattagta atctaaaagc
     2221 actccgataa cgattagagt gcttttagat tgtttatcat taattatcaa agcaagtgca
     2281 gttcattatg ccaggatgcg ctcgcacaga cgccaacccg tgacaccgtt gatggtcacg
     2341 cggaacagca ggctgccgtc cgggttaatc agacgctcgt caataatctt attgccattc
     2401 cacagggtac cggtgacggt gatcttttta ccgtcaaaaa cggcaatgcc ttcatacgga
     2461 cggccgaaat agtcaatcat attcggggtc acaccatcaa tcaccagcgt accgtaatgc
     2521 aagataacct tgaagtggtg atcgtcaact gggtagacga ctttaaagat tttctcgatt
     2581 tgacccattt ggtcaccgct caggccttcg tacgggatga tcacatgaat atcaattttc
     2641 agaccgttct cgccggacag aacgatgcgc tgaatcggcg tcacgctcac acccaggttc
     2701 tggaacagag agcttacgcc accctgttcc aggacttgat ccaaattata acccgcggtc
     2761 tgacgccaat cgccaacaaa atcttccaga gtaaaaacca tgtttatatt atttatattt
     2821 gtttgacgag aatatcttta tttgccgaca aaggtacata actaaagttt cccacccaaa
     2881 taaatagata gaaaaataac agtttgtcga attttctttg tacccgatac ttgtgcctgt
     2941 tctatttccg aaccgaccgc ttgtatgaat gcatcaaaat tcgttttctc tacgttggat
     3001 tccttgttgc tcatattgtg atgataattt ctacaaatat agtcattggt aactatctat
     3061 gaaactgttt gatactttta tagttgatta aacttgttca tggcatttgc cttaatatca
     3121 tccgctatgt caatgtaggg tttcatagct ttgtagtcgc tgtgtcccgt ccatttcatg
     3181 accacctgtg ccgggattcc gagagccagc gcattgcaga tgaatgtcct ttttcctgca
     3241 tgggtactga gcaaagcgta tttgggtgtg acttcatcaa tacgttcatt tcccttgtag
     3301 taggtttccc gtacaggctc gttgatttct gccagttcac ccagctcttt caggtaatcg
     3361 ttcatcttct ggttgctgat gacgggcaga gccatgtaat tctcgaaatg gatgtccttg
     3421 tatttgtcca gtatggcttt gctgtatttg ttcagttcaa tcgtcaggct gtcggcagtc
     3481 ttgactgtgg ttatttcgat gtggtcggac ttcacatcgc ttcttttcag attgcgaaca
     3541 tccgaatacc gcaaactcgt aaagcagcag aacaggaaaa catcacgcac acgttccagg
     3601 tattgcttat ccttgggtat ctggtagtct ttcagcttgt tcagttcatc ccaagtcagg
     3661 aagattactt ttttcgaggt ggttttcagt ttcggtttga acgtatcgta tgcaatgttc
     3721 tgatgatgtc ctttcttgaa gctccagcgc aggaaccatt tgaggaatcc catttgcttg
     3781 ccgatggtgc tgtttctcat atccttggtg tcacgcagga agttgacgta ttcgttcaat
     3841 ccaaactcgt tgaaatagtt gaacgttgca tcctccttga actctttgag gtggttcctc
     3901 actgctgcaa atttttcata ggtggatgcc gtccagttat tctggttacc gcactctttt
     3961 acaaactcat cgaacacctc ccaaaagctg acaggggctt cttccggctg ttcttcactg
     4021 gtgtctttca ttctcatgtt gaaagcttcc ttcaactgtt gggtcgttgg catgacctcc
     4081 tgcacctcaa attccttgaa aatattctgg atttcggcat agtatttcag caagtccgta
     4141 ttgatttcgg ctgcactttg ctttagcttg ttggtacatc cgttctttac ccgctgctta
     4201 tctacatccc atttggctac gtcaatccgg tagcccgttg taaactcgat acgttggctg
     4261 gcaaagatga cacgcatacg gatgggtacg ttctctacga ttggcacacc gttctttttc
     4321 cggctctcca atgcaaaaat gatgttgcgc ttgatattca taattgggtg cgtttgaaat
     4381 tctacaccca aatatacacc caattattga gatagcaaaa gacatttaga aacatttact
     4441 tttactctat attgtaattt acacttgatt atcagtcgtt tgcagtttta tgatattctg
     4501 tgaaagtata agttcgagag cctgtctctc cgcaaaaaac gctgaaaatc agcagattgc
     4561 aaaacaaaca ccctgtttta cacccaagaa tgtaaagtcg ggtgtttttg ttttatttaa
     4621 gataatacaa ccactacata ataaaagagt agcgatatta aaagaatccg atgagaaaag
     4681 actaatattt atctatccat tcagtttgat ttctcaggac tttacatcgt cctgaaagta
     4741 tttgttggta ccgaggacgc gtaaacattt acagttgcat gtggcctatt gtttttagcc
     4801 gttaaatatt ttataactat taaatagcga tacaaattgt tcgaaactaa tattgtttat
     4861 atcatatatt ctcgcatgtt ttaaagcttt attaaattga ttttttgtaa acagtttttc
     4921 gtactctttg ttaacccatt tcattacaaa agtttcatat ttttttctct ctttaaatgc
     4981 catttttgct ggctttcttt ttaatacaat taatgtgcta tccactttag gttttggatg
     5041 gaaataatac ctaggaattt ttgctaatat agaaatatct acctctgcca ttaacagcaa
     5101 tgctagtgat ctgtttgtat ctaataacat tttagcaaaa ccatattcca ctattaaata
     5161 acttattgtg gctgaacttt caaaaacaat ttttcgaatt atatttgtgc ttatgttgta
     5221 aggtatgctg ccaaatattt tatatggatt gtggctagga aatgtaaatt tcagtatatc
     5281 atcatttact atttgatagt taggataatt taagagctta ttacgagtta cctcacataa
     5341 tttagaatca atttctatcg ccgttacaaa attacatctc tttaccaatc cagcagtaaa
     5401 atgacctttc cctgcaccta tttcaaagat gttatctttt tcatctaaac ttatgcaatt
     5461 cattattttt tctatgtgat attttgaagt aataaaattt tgactatctt ttatatttac
     5521 tttgttcatt ataacctctc cttaatttat tgcatctctt ttcgaatatt tatgtttttt
     5581 gagaaaagaa cgtactcatg gttcatcccg atatgcgtat cggtctgtat atcagcaact
     5641 ttctatgtgt ttcaactaca atagtcatct attctcatct ttctgagtcc accccctgca
     5701 aagcccctct ttacgacata aaaattcggt cggaaaaggt atgcaaaaga tgtttctctc
     5761 tttaagagaa actcttcggg atgcaaaaat atgaaaataa ctccaattca ccaaattata
     5821 tagcgacttt tttacaaaat gctaaaattt gttgatttcc gtcaagcaat tgttgagcaa
     5881 aaatgtcttt tacgataaaa tgatacctca atatcaactg tttagcaaaa cgatatttct
     5941 cttaaagaga gaaacacctt tttgttcacc aatccccgac ttttaatccc gcggccatga
     6001 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     6061 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     6121 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     6181 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     6241 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     6301 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     6361 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     6421 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     6481 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6541 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     6601 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     6661 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6721 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6781 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6841 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     6901 acgcgtcaat tcgaggggga tcaattccgt gataggtggg ctgcccttcc tggttggctt
     6961 ggtttcatca gccatccgct tgccctcatc tgttacgccg gcggtagccg gccagcctcg
     7021 cagagcagga ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca
     7081 cccgctcgcg ggtgggccta cttcacctat cctgcccggc tgacgccgtt ggatacacca
     7141 aggaaagtct acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata
     7201 taccgaaaaa atcgctataa tgaccccgaa gcagggttat gcagcggaaa acggaattga
     7261 tccggccacg atgcgtccgg cgtagaggat ctgaagatca gcagttcaac ctgttgatag
     7321 tacgtactaa gctctcatgt ttcacgtact aagctctcat gtttaacgta ctaagctctc
     7381 atgtttaacg aactaaaccc tcatggctaa cgtactaagc tctcatggct aacgtactaa
     7441 gctctcatgt ttcacgtact aagctctcat gtttgaacaa taaaattaat ataaatcagc
     7501 aacttaaata gcctctaagg ttttaagttt tataagaaaa aaaagaatat ataaggcttt
     7561 taaagctttt aag
//
