LOCUS       sequence-197114-        5785 bp    DNA     linear   UNA 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 469.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5785)
  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 5785)
  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: MK1282
FEATURES             Location/Qualifiers
     source          1..5785
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(62..80)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     128..146
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        577..595
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             684..704
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      3373..3444
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3460..3487
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(3549..3565)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3550..3566)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(3594..3612)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3633..3649)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3633..3649)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3646..3668)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3657..3673
                     /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(3681..3687,3688..3705,3706..3711))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3726..3747
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3864..3881)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4035..4623)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4115..4134)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4794..5585,5586..5654))
                     /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     5417..5436
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5655..5759)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atggtggtag acatttttta ccgcccgtag aattcggtgg gtgctggtgc cccgggatag
       61 ccctatagtg agtcgtatta atttccgcgg gcgagctccc attattatca tgacattaac
      121 ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattgatct gtcgacagga
      181 actggacagc gtgtcgaaaa agctcgacag gaactggaca gcgtgtcgaa aaagctcgac
      241 aggaactgga cagcgtgtcg aaaaagctcg acaggaactg gacagcgtgt cgaaaaagct
      301 cgagcagatc cgccaggcgt gtatatatag cgtggatggc caggcaactt tagtgctgac
      361 acatacaggc atatatatat gtgtgcgacg acacatgatc atatggcatg catgtgctct
      421 gtatgtatat aaaactcttg ttttcttctt ttctctaaat attctttcct tatacattag
      481 gtcctttgta gcataaatta ctatacttct atagacacgc aaacacaaat acacacacta
      541 aattaataac tagttctatc attgataggt ataaattaat acgactcact atagggagac
      601 ctatcagtga tagaggatcc aaccgagccc aatataggac tcagggtgcc aaaaaaatgt
      661 ctaccaccat gggaattcaa cctcctaaaa agaaacgtaa agttaatact attaatattg
      721 ctaaaaatga cttctcagat attgaattag cagccattcc atttaataca ttagcagatc
      781 actatggtga acgtttagca cgtgaacagt tagcattaga acatgaatca tatgaaatgg
      841 gtgaagcacg ttttcgtaag atgttcgagc gtcagttaaa agcaggtgaa gttgcagata
      901 atgcagcagc caaaccttta attactacat tattacctaa aatgattgct cgtattaacg
      961 attggtttga agaggttaaa gcaaagcgtg gtaaacgtcc tacagcattt cagttcttac
     1021 aagaaatcaa acctgaagca gttgcatata ttactattaa aacaacatta gcatgtttaa
     1081 catcagcaga taatacaaca gttcaagcag ttgcatcagc aattggtcgt gcaattgaag
     1141 atgaagcacg ttttggtcgt attcgtgatt tagaagccaa acattttaaa aaaaatgttg
     1201 aagaacagtt aaacaaacgt gttggtcatg tttataaaaa agcatttatg caggttgttg
     1261 aagcagatat gttatcaaaa ggtttattag gtggtgaagc atggtcatca tggcataaag
     1321 aagattcaat tcatgttggt gttcgttgta ttgaaatgtt aattgaatca acaggtatgg
     1381 tttcattaca tcgtcagaat gcaggtgttg ttggtcaaga ttcagaaaca attgaattag
     1441 cacctgaata tgcagaagca attgcaacac gtgcaggtgc attagcaggt atttcaccaa
     1501 tgtttcagcc ttgtgttgtt cctcctaaac cttggacagg tattacaggt ggtggttatt
     1561 gggcaaatgg tcgtcgtcct ttagcattag ttcgtacaca ttcaaaaaaa gcattaatgc
     1621 gttatgaaga tgtttacatg cctgaagttt ataaagccat taatattgca cagaatactg
     1681 catggaaaat caacaagaaa gttttagcag ttgcaaatgt tattactaaa tggaaacatt
     1741 gtcctgttga agatattcct gcaattgaac gtgaagaatt accaatgaaa cctgaagata
     1801 ttgatatgaa tcctgaagca ttaacagcat ggaaacgtgc agcagcagcc gtttatcgta
     1861 aagataaagc acgtaaatca cgtcgtattt cattagagtt catgttagag caagcaaaca
     1921 agttcgcaaa tcataaagcc atttggtttc cttataatat ggattggcgt ggtcgtgttt
     1981 atgcagtttc aatgtttaat cctcaaggta atgatatgac caaaggttta ttaaccttag
     2041 ctaaaggtaa acctattggt aaagaaggtt attattggtt aaaaatccat ggtgcaaatt
     2101 gtgcaggtgt tgataaagtt ccttttccag aacgtattaa gttcattgaa gaaaatcatg
     2161 aaaatattat ggcatgtgct aaatcaccat tagaaaatac atggtgggca gaacaagatt
     2221 cacctttttg ttttttagcc ttttgttttg aatatgcagg tgttcaacat catggtttat
     2281 catataattg ttcattacca ttagcatttg atggttcatg ttcaggtatt cagcattttt
     2341 cagcaatgtt acgtgatgaa gttggtggtc gtgcagttaa cttattacct tcagaaacag
     2401 ttcaagatat ctatggtatt gttgctaaaa aagttaatga aatcttacag gcagatgcca
     2461 ttaatggtac agataatgaa gttgttacag ttacagatga aaatacaggt gaaatatcag
     2521 aaaaagttaa attaggtacc aaagcattag caggtcaatg gttagcatat ggtgttacac
     2581 gttcagttac taagcgttca gttatgacat tagcatatgg ttcaaaagag ttcggttttc
     2641 gtcaacaggt tttagaagat accattcaac ctgcaattga ttcaggtaaa ggtttaatgt
     2701 ttactcaacc taatcaagca gcaggttata tggccaaatt aatatgggaa tcagtttcag
     2761 ttacagttgt tgcagcagtt gaagcaatga attggttaaa atcagcagcc aaattattag
     2821 cagcagaagt taaagataaa aaaacaggtg aaatcttacg taagcgttgt gcagttcatt
     2881 gggttacacc tgatggtttt cctgtttggc aagaatataa aaaacctatt caaacacgtt
     2941 taaacttaat gtttttaggt cagttccgtt tacaacctac tattaataca aacaaagatt
     3001 cagaaattga tgcacataaa caagaatcag gtattgcacc taacttcgtt cattcacaag
     3061 atggttcaca tttacgtaaa acagttgttt gggcacatga aaaatatggt attgaatcat
     3121 ttgcattaat tcacgattca tttggtacca ttccagcaga tgcagcaaac ttattcaaag
     3181 cagttcgtga aacaatggtt gatacatatg aatcatgtga tgttttagca gacttctatg
     3241 atcagttcgc agatcagtta catgaatcac agttagataa aatgcctgca ttacctgcca
     3301 aaggtaactt aaacttacgt gatattttag aatcagactt cgcatttgcc taataaggcg
     3361 cgccccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     3421 tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct
     3481 ttctgcgttt atagctatta ctcgaacccc tagcccgctc ttatcgggcg gctaggggtt
     3541 ttttgtatcg ataccgtcga cctcgagggg gggcccggta ccagcttttg ttccctttag
     3601 tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     3661 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3721 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3781 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3841 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3901 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3961 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4021 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4081 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4141 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4201 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4261 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4321 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4381 gcagcagcaa ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4441 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4501 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4561 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4621 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4681 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4741 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4801 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4861 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4921 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4981 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5041 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5101 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5161 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5221 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5281 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5341 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5401 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5461 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5521 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5581 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5641 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttagca gggttattgt
     5701 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5761 acatttcccc gaaaagtgcc acctg
//
