LOCUS       sequence-197095-        7183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This E. coli DH10B strain harbors a shuttle vector plasmid that 
            expresses the Mixed Feedback Loop version of the UBER system with a 
            T7RNAP translation rate of 300 and a TetR translation rate of 
            35149..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7183)
  AUTHORS   Kushwaha M, Salis HM
  TITLE     A portable expression resource for engineering cross-species genetic
            circuits and pathways.
  JOURNAL   Nat Commun. 2015 Jul 17;6:7832. doi: 10.1038/ncomms8832.
  PUBMED    26184393
REFERENCE   2  (bases 1 to 7183)
  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: "2015-07-17"; volume: "6:7832. doi"; pages: " 
            10.1038/ncomms8832"
COMMENT     Sequence Label: MK1274
FEATURES             Location/Qualifiers
     source          1..7183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(182..203)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     394..413
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(1383..1403)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(1478..1496)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1544..1562
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1993..2011
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2100..2120
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      4789..4860
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4876..4903
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(4965..4981)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4966..4982)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(5010..5028)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5049..5065)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5049..5065)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5062..5084)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5073..5089
                     /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)."
     promoter        complement(join(5097..5103,5104..5121,5122..5127))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5142..5163
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5280..5297)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5451..6039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5531..5550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6210..7001,7002..7070))
                     /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     6833..6852
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7071..7175)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgaaatacc gcacagatgc gtaaggagaa gataccgcay caggaaattg taagcgttaa
       61 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
      121 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
      181 tccagtttgg aacaagagtc cactattaat gaatgtattt agaaaaataa acaaataggg
      241 gttccgcgca catttcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
      301 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
      361 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
      421 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
      481 tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
      541 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
      601 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaacgacg gccagtgaat
      661 tggcgaattg gagctattaa ttaaacaaaa aacccctacc cccccgatat gtatttattg
      721 cgaaaaaacc ccgccgaagc ggggtttttt gcgaccggtt attatgaacc tgattcacat
      781 tttaactgtt tctccaaacc gcaaatgatt aattctaaac caaataaaaa tgcaggttct
      841 gcaccttgat gatcaaataa ttcaattgct tgacgtaata aaggaggcat tgaatctgtt
      901 gtaggtgttt ctctttcctc cttggcaact tgatgttctt gatcttctaa aacacaacct
      961 aaggtaaaat gaccaactgc tgataatgca tataatgcgt tttctaatga aaaaccttgt
     1021 tgacataaga atgctaactg attttctaat gtttcatatt gtttttctgt aggacgtgta
     1081 cctaagtgaa ctttggcacc atctctatgt gataataatg cacaacgaaa tgatttggca
     1141 ttgttacgta agaagtcttg ccatgattct ccttctaatg gacaaaaatg tgtatgatgt
     1201 ctatctaaca tttcgattgc taatgcatct aataatgcac gtttgttttt aacatgccaa
     1261 tataatgtag gttgttcaac acctaatttc tgtgctaatt tacgtgtagt taaaccttcg
     1321 ataccaactt catttaataa ttctaatgct gagttaataa cttttgattt atctaaacgt
     1381 gaaactttac gttttttttt aggaggatag atctccatgg tggtagacat ttttgaccgc
     1441 ccttagaatt cggcgcgtgc tggtgccccg ggatagccct atagtgagtc gtattaattt
     1501 ccgcgggcga gctcccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     1561 gccctttcgt cttcaagaat tgatctgtcg acaggaactg gacagcgtgt cgaaaaagct
     1621 cgacaggaac tggacagcgt gtcgaaaaag ctcgacagga actggacagc gtgtcgaaaa
     1681 agctcgacag gaactggaca gcgtgtcgaa aaagctcgag cagatccgcc aggcgtgtat
     1741 atatagcgtg gatggccagg caactttagt gctgacacat acaggcatat atatatgtgt
     1801 gcgacgacac atgatcatat ggcatgcatg tgctctgtat gtatataaaa ctcttgtttt
     1861 cttcttttct ctaaatattc tttccttata cattaggtcc tttgtagcat aaattactat
     1921 acttctatag acacgcaaac acaaatacac acactaaatt aataactagt tctatcattg
     1981 ataggtataa attaatacga ctcactatag ggagacctat cagtgataga ggatccaacc
     2041 gagcccaata taggactcag ggtgccaaaa aaatgtctac caccatggga attcaacctc
     2101 ctaaaaagaa acgtaaagtt aatactatta atattgctaa aaatgacttc tcagatattg
     2161 aattagcagc cattccattt aatacattag cagatcacta tggtgaacgt ttagcacgtg
     2221 aacagttagc attagaacat gaatcatatg aaatgggtga agcacgtttt cgtaagatgt
     2281 tcgagcgtca gttaaaagca ggtgaagttg cagataatgc agcagccaaa cctttaatta
     2341 ctacattatt acctaaaatg attgctcgta ttaacgattg gtttgaagag gttaaagcaa
     2401 agcgtggtaa acgtcctaca gcatttcagt tcttacaaga aatcaaacct gaagcagttg
     2461 catatattac tattaaaaca acattagcat gtttaacatc agcagataat acaacagttc
     2521 aagcagttgc atcagcaatt ggtcgtgcaa ttgaagatga agcacgtttt ggtcgtattc
     2581 gtgatttaga agccaaacat tttaaaaaaa atgttgaaga acagttaaac aaacgtgttg
     2641 gtcatgttta taaaaaagca tttatgcagg ttgttgaagc agatatgtta tcaaaaggtt
     2701 tattaggtgg tgaagcatgg tcatcatggc ataaagaaga ttcaattcat gttggtgttc
     2761 gttgtattga aatgttaatt gaatcaacag gtatggtttc attacatcgt cagaatgcag
     2821 gtgttgttgg tcaagattca gaaacaattg aattagcacc tgaatatgca gaagcaattg
     2881 caacacgtgc aggtgcatta gcaggtattt caccaatgtt tcagccttgt gttgttcctc
     2941 ctaaaccttg gacaggtatt acaggtggtg gttattgggc aaatggtcgt cgtcctttag
     3001 cattagttcg tacacattca aaaaaagcat taatgcgtta tgaagatgtt tacatgcctg
     3061 aagtttataa agccattaat attgcacaga atactgcatg gaaaatcaac aagaaagttt
     3121 tagcagttgc aaatgttatt actaaatgga aacattgtcc tgttgaagat attcctgcaa
     3181 ttgaacgtga agaattacca atgaaacctg aagatattga tatgaatcct gaagcattaa
     3241 cagcatggaa acgtgcagca gcagccgttt atcgtaaaga taaagcacgt aaatcacgtc
     3301 gtatttcatt agagttcatg ttagagcaag caaacaagtt cgcaaatcat aaagccattt
     3361 ggtttcctta taatatggat tggcgtggtc gtgtttatgc agtttcaatg tttaatcctc
     3421 aaggtaatga tatgaccaaa ggtttattaa ccttagctaa aggtaaacct attggtaaag
     3481 aaggttatta ttggttaaaa atccatggtg caaattgtgc aggtgttgat aaagttcctt
     3541 ttccagaacg tattaagttc attgaagaaa atcatgaaaa tattatggca tgtgctaaat
     3601 caccattaga aaatacatgg tgggcagaac aagattcacc tttttgtttt ttagcctttt
     3661 gttttgaata tgcaggtgtt caacatcatg gtttatcata taattgttca ttaccattag
     3721 catttgatgg ttcatgttca ggtattcagc atttttcagc aatgttacgt gatgaagttg
     3781 gtggtcgtgc agttaactta ttaccttcag aaacagttca agatatctat ggtattgttg
     3841 ctaaaaaagt taatgaaatc ttacaggcag atgccattaa tggtacagat aatgaagttg
     3901 ttacagttac agatgaaaat acaggtgaaa tatcagaaaa agttaaatta ggtaccaaag
     3961 cattagcagg tcaatggtta gcatatggtg ttacacgttc agttactaag cgttcagtta
     4021 tgacattagc atatggttca aaagagttcg gttttcgtca acaggtttta gaagatacca
     4081 ttcaacctgc aattgattca ggtaaaggtt taatgtttac tcaacctaat caagcagcag
     4141 gttatatggc caaattaata tgggaatcag tttcagttac agttgttgca gcagttgaag
     4201 caatgaattg gttaaaatca gcagccaaat tattagcagc agaagttaaa gataaaaaaa
     4261 caggtgaaat cttacgtaag cgttgtgcag ttcattgggt tacacctgat ggttttcctg
     4321 tttggcaaga atataaaaaa cctattcaaa cacgtttaaa cttaatgttt ttaggtcagt
     4381 tccgtttaca acctactatt aatacaaaca aagattcaga aattgatgca cataaacaag
     4441 aatcaggtat tgcacctaac ttcgttcatt cacaagatgg ttcacattta cgtaaaacag
     4501 ttgtttgggc acatgaaaaa tatggtattg aatcatttgc attaattcac gattcatttg
     4561 gtaccattcc agcagatgca gcaaacttat tcaaagcagt tcgtgaaaca atggttgata
     4621 catatgaatc atgtgatgtt ttagcagact tctatgatca gttcgcagat cagttacatg
     4681 aatcacagtt agataaaatg cctgcattac ctgccaaagg taacttaaac ttacgtgata
     4741 ttttagaatc agacttcgca tttgcctaat aaggcgcgcc ccaggcatca aataaaacga
     4801 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     4861 tactagagtc acactggctc accttcgggt gggcctttct gcgtttatag ctattactcg
     4921 aacccctagc ccgctcttat cgggcggcta ggggtttttt gtatcgatac cgtcgacctc
     4981 gagggggggc ccggtaccag cttttgttcc ctttagtgag ggttaatttc gagcttggcg
     5041 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5101 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5161 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5221 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5281 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5341 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5401 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5461 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5521 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5581 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5641 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5701 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5761 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagcaactgg taacaggatt
     5821 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5881 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5941 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6001 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6061 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6121 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6181 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6241 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6301 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6361 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6421 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6481 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6541 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6601 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6661 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6721 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6781 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6841 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6901 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6961 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7021 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7081 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7141 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttc
//
