LOCUS       sequence-181560-        7773 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 1984 and a TetR translation rate of 
            3877..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7773)
  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 7773)
  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: MK1278
FEATURES             Location/Qualifiers
     source          1..7773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(65..169)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(295..317)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     417..436
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        complement(470..574)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(600..1055)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(737..758)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     949..968
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1182..1204
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1196..1213
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1197..1213
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1749..1769)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(1844..1862)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2359..2377
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2466..2486
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5155..5226
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5242..5269
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(5331..5347)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5332..5348)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(5376..5394)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5415..5431)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5415..5431)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5428..5450)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5439..5455
                     /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(5463..5469,5470..5487,5488..5493))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5508..5529
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5646..5663)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5817..6405)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5897..5916)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6576..7367,7368..7436))
                     /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     7199..7218
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7437..7541)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
       61 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
      121 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
      181 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
      241 ggcgtatcac gaggcccttt cgtctcgcgc gcgtttcggt gatgacggtg aaaacctctg
      301 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
      361 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
      421 atcagagcag attgtactga gagtgcacac ggaaatgttg aatactcata ctcttccttt
      481 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
      541 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
      601 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
      661 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
      721 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
      781 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
      841 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
      901 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
      961 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     1021 acccgccgcg cttaatgcgc cgctacaggg cgcgtccnat tcgccattca ggctgcgcaa
     1081 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     1141 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     1201 aacgacggcc agtgaattgt aatacgactc actatarkga aayggataca tatttgaatg
     1261 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg
     1321 gtccttttca tcacgtgcta taaaaataat tataatttaa attcgttttc taatgaaaaa
     1381 ccttgttgac ataagaatgc taactgattt tctaatgttt catattgttt ttctgtagga
     1441 cgtgtaccta agtgaacttt ggcaccatct ctatgtgata ataatgcaca acgaaatgat
     1501 ttggcattgt tacgtaagaa gtcttgccat gattctcctt ctaatggaca aaaatgtgta
     1561 tgatgtctat ctaacatttc gattgctaat gcatctaata atgcacgttt gtttttaaca
     1621 tgccaatata atgtaggttg ttcaacacct aatttctgtg ctaatttacg tgtagttaaa
     1681 ccttcgatac caacttcatt taataattct aatgctgagt taataacttt tgatttatct
     1741 aaacgtgaaa ctttacgttt ttttttagga ggatagatct ccatggtggt agacattttt
     1801 ttggctccat agrwttcggc gggtgctggt gccccgggat agccctatag tgagtcgtat
     1861 taatttccgc gggcgagctc ccattattat catgacatta acctataaaa ataggcgtat
     1921 cacgaggccc tttcgtcttc aagaattgat ctgtcgacag gaactggaca gcgtgtcgaa
     1981 aaagctcgac aggaactgga cagcgtgtcg aaaaagctcg acaggaactg gacagcgtgt
     2041 cgaaaaagct cgacaggaac tggacagcgt gtcgaaaaag ctcgagcaga tccgccaggc
     2101 gtgtatatat agcgtggatg gccaggcaac tttagtgctg acacatacag gcatatatat
     2161 atgtgtgcga cgacacatga tcatatggca tgcatgtgct ctgtatgtat ataaaactct
     2221 tgttttcttc ttttctctaa atattctttc cttatacatt aggtcctttg tagcataaat
     2281 tactatactt ctatagacac gcaaacacaa atacacacac taaattaata actagttcta
     2341 tcattgatag gtataaatta atacgactca ctatagggag acctatcagt gatagaggat
     2401 ccaaccgagc ccaatatagg acctatggag sggcaaaaat gtctaccacc atgggaattc
     2461 aacctcctaa aaagaaacgt aaagttaata ctattaatat tgctaaaaat gacttctcag
     2521 atattgaatt agcagccatt ccatttaata cattagcaga tcactatggt gaacgtttag
     2581 cacgtgaaca gttagcatta gaacatgaat catatgaaat gggtgaagca cgttttcgta
     2641 agatgttcga gcgtcagtta aaagcaggtg aagttgcaga taatgcagca gccaaacctt
     2701 taattactac attattacct aaaatgattg ctcgtattaa cgattggttt gaagaggtta
     2761 aagcaaagcg tggtaaacgt cctacagcat ttcagttctt acaagaaatc aaacctgaag
     2821 cagttgcata tattactatt aaaacaacat tagcatgttt aacatcagca gataatacaa
     2881 cagttcaagc agttgcatca gcaattggtc gtgcaattga agatgaagca cgttttggtc
     2941 gtattcgtga tttagaagcc aaacatttta aaaaaaatgt tgaagaacag ttaaacaaac
     3001 gtgttggtca tgtttataaa aaagcattta tgcaggttgt tgaagcagat atgttatcaa
     3061 aaggtttatt aggtggtgaa gcatggtcat catggcataa agaagattca attcatgttg
     3121 gtgttcgttg tattgaaatg ttaattgaat caacaggtat ggtttcatta catcgtcaga
     3181 atgcaggtgt tgttggtcaa gattcagaaa caattgaatt agcacctgaa tatgcagaag
     3241 caattgcaac acgtgcaggt gcattagcag gtatttcacc aatgtttcag ccttgtgttg
     3301 ttcctcctaa accttggaca ggtattacag gtggtggtta ttgggcaaat ggtcgtcgtc
     3361 ctttagcatt agttcgtaca cattcaaaaa aagcattaat gcgttatgaa gatgtttaca
     3421 tgcctgaagt ttataaagcc attaatattg cacagaatac tgcatggaaa atcaacaaga
     3481 aagttttagc agttgcaaat gttattacta aatggaaaca ttgtcctgtt gaagatattc
     3541 ctgcaattga acgtgaagaa ttaccaatga aacctgaaga tattgatatg aatcctgaag
     3601 cattaacagc atggaaacgt gcagcagcag ccgtttatcg taaagataaa gcacgtaaat
     3661 cacgtcgtat ttcattagag ttcatgttag agcaagcaaa caagttcgca aatcataaag
     3721 ccatttggtt tccttataat atggattggc gtggtcgtgt ttatgcagtt tcaatgttta
     3781 atcctcaagg taatgatatg accaaaggtt tattaacctt agctaaaggt aaacctattg
     3841 gtaaagaagg ttattattgg ttaaaaatcc atggtgcaaa ttgtgcaggt gttgataaag
     3901 ttccttttcc agaacgtatt aagttcattg aagaaaatca tgaaaatatt atggcatgtg
     3961 ctaaatcacc attagaaaat acatggtggg cagaacaaga ttcacctttt tgttttttag
     4021 ccttttgttt tgaatatgca ggtgttcaac atcatggttt atcatataat tgttcattac
     4081 cattagcatt tgatggttca tgttcaggta ttcagcattt ttcagcaatg ttacgtgatg
     4141 aagttggtgg tcgtgcagtt aacttattac cttcagaaac agttcaagat atctatggta
     4201 ttgttgctaa aaaagttaat gaaatcttac aggcagatgc cattaatggt acagataatg
     4261 aagttgttac agttacagat gaaaatacag gtgaaatatc agaaaaagtt aaattaggta
     4321 ccaaagcatt agcaggtcaa tggttagcat atggtgttac acgttcagtt actaagcgtt
     4381 cagttatgac attagcatat ggttcaaaag agttcggttt tcgtcaacag gttttagaag
     4441 ataccattca acctgcaatt gattcaggta aaggtttaat gtttactcaa cctaatcaag
     4501 cagcaggtta tatggccaaa ttaatatggg aatcagtttc agttacagtt gttgcagcag
     4561 ttgaagcaat gaattggtta aaatcagcag ccaaattatt agcagcagaa gttaaagata
     4621 aaaaaacagg tgaaatctta cgtaagcgtt gtgcagttca ttgggttaca cctgatggtt
     4681 ttcctgtttg gcaagaatat aaaaaaccta ttcaaacacg tttaaactta atgtttttag
     4741 gtcagttccg tttacaacct actattaata caaacaaaga ttcagaaatt gatgcacata
     4801 aacaagaatc aggtattgca cctaacttcg ttcattcaca agatggttca catttacgta
     4861 aaacagttgt ttgggcacat gaaaaatatg gtattgaatc atttgcatta attcacgatt
     4921 catttggtac cattccagca gatgcagcaa acttattcaa agcagttcgt gaaacaatgg
     4981 ttgatacata tgaatcatgt gatgttttag cagacttcta tgatcagttc gcagatcagt
     5041 tacatgaatc acagttagat aaaatgcctg cattacctgc caaaggtaac ttaaacttac
     5101 gtgatatttt agaatcagac ttcgcatttg cctaataagg cgcgccccag gcatcaaata
     5161 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     5221 gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttatagctat
     5281 tactcgaacc cctagcccgc tcttatcggg cggctagggg ttttttgtat cgataccgtc
     5341 gacctcgagg gggggcccgg taccagcttt tgttcccttt agtgagggtt aatttcgagc
     5401 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5461 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5521 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5581 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5641 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5701 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5761 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5821 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5881 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5941 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6001 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6061 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6121 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc aactggtaac
     6181 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6241 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6301 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6361 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6421 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6481 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6541 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6601 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6661 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6721 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6781 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6841 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6901 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6961 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7021 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7081 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7141 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7201 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7261 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7321 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7381 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7441 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7501 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7561 ccacctgggt ccttttcatc acgtgctata aaaataatta taatttaaat tcgttttcta
     7621 atgaaaaacc ttgttgacat aagaatgcta actgattttc taatgtttca tattgttttt
     7681 ctgtaggacg tgtacctaag tgaactttgg caccatctct atgtgataat aatgcacaac
     7741 gaaatgattt ggcattgtta cgtaagaaga gtc
//
