LOCUS       sequence-440608-        6436 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 1535 and a TetR translation rate of 
            30709.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6436)
  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 6436)
  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: MK1283
FEATURES             Location/Qualifiers
     source          1..6436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(936..956)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(1031..1049)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1097..1115
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1546..1564
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1653..1673
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      4342..4413
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4429..4456
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(4518..4534)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4519..4535)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(4563..4581)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4602..4618)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4602..4618)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4615..4637)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4626..4642
                     /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(4650..4656,4657..4674,4675..4680))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4695..4716
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4833..4850)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5004..5592)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5084..5103)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5763..6436,1..118,119..187))
                     /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     6386..6405
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
       61 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
      121 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
      181 tactcatact cttccaaacg acggccagtg aattggcgaa ttggagctat taattaaaca
      241 aaaaacccct acccccccga tatgtattta ttgcgaaaaa accccgccga agcggggttt
      301 tttgcgaccg gttattatga acctgattca cattttaact gtttctccaa accgcaaatg
      361 attaattcta aaccaaataa aaatgcaggt tctgcacctt gatgatcaaa taattcaatt
      421 gcttgacgta ataaaggagg cattgaatct gttgtaggtg tttctctttc ctccttggca
      481 acttgatgtt cttgatcttc taaaacacaa cctaaggtaa aatgaccaac tgctgataat
      541 gcatataatg cgttttctaa tgaaaaacct tgttgacata agaatgctaa ctgattttct
      601 aatgtttcat attgtttttc tgtaggacgt gtacctaagt gaactttggc accatctcta
      661 tgtgataata atgcacaacg aaatgatttg gcattgttac gtaagaagtc ttgccatgat
      721 tctccttcta atggacaaaa atgtgtatga tgtctatcta acatttcgat tgctaatgca
      781 tctaataatg cacgtttgtt tttaacatgc caatataatg taggttgttc aacacctaat
      841 ttctgtgcta atttacgtgt agttaaacct tcgataccaa cttcatttaa taattctaat
      901 gctgagttaa taacttttga tttatctaaa cgtgaaactt tacgtttttt tttaggagga
      961 tagatctcca tggtggtaga catttttgac cgcccttaga attcggtgcg tgctggtgcc
     1021 ccgggatagc cctatagtga gtcgtattaa tttccgcggg cgagctccca ttattatcat
     1081 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag aattgatctg
     1141 tcgacaggaa ctggacagcg tgtcgaaaaa gctcgacagg aactggacag cgtgtcgaaa
     1201 aagctcgaca ggaactggac agcgtgtcga aaaagctcga caggaactgg acagcgtgtc
     1261 gaaaaagctc gagcagatcc gccaggcgtg tatatatagc gtggatggcc aggcaacttt
     1321 agtgctgaca catacaggca tatatatatg tgtgcgacga cacatgatca tatggcatgc
     1381 atgtgctctg tatgtatata aaactcttgt tttcttcttt tctctaaata ttctttcctt
     1441 atacattagg tcctttgtag cataaattac tatacttcta tagacacgca aacacaaata
     1501 cacacactaa attaataact agttctatca ttgataggta taaattaata cgactcacta
     1561 tagggagacc tatcagtgat agaggatcca accgagccca atataggacc tagggtgcca
     1621 aaaaaatgtc taccaccatg ggaattcaac ctcctaaaaa gaaacgtaaa gttaatacta
     1681 ttaatattgc taaaaatgac ttctcagata ttgaattagc agccattcca tttaatacat
     1741 tagcagatca ctatggtgaa cgtttagcac gtgaacagtt agcattagaa catgaatcat
     1801 atgaaatggg tgaagcacgt tttcgtaaga tgttcgagcg tcagttaaaa gcaggtgaag
     1861 ttgcagataa tgcagcagcc aaacctttaa ttactacatt attacctaaa atgattgctc
     1921 gtattaacga ttggtttgaa gaggttaaag caaagcgtgg taaacgtcct acagcatttc
     1981 agttcttaca agaaatcaaa cctgaagcag ttgcatatat tactattaaa acaacattag
     2041 catgtttaac atcagcagat aatacaacag ttcaagcagt tgcatcagca attggtcgtg
     2101 caattgaaga tgaagcacgt tttggtcgta ttcgtgattt agaagccaaa cattttaaaa
     2161 aaaatgttga agaacagtta aacaaacgtg ttggtcatgt ttataaaaaa gcatttatgc
     2221 aggttgttga agcagatatg ttatcaaaag gtttattagg tggtgaagca tggtcatcat
     2281 ggcataaaga agattcaatt catgttggtg ttcgttgtat tgaaatgtta attgaatcaa
     2341 caggtatggt ttcattacat cgtcagaatg caggtgttgt tggtcaagat tcagaaacaa
     2401 ttgaattagc acctgaatat gcagaagcaa ttgcaacacg tgcaggtgca ttagcaggta
     2461 tttcaccaat gtttcagcct tgtgttgttc ctcctaaacc ttggacaggt attacaggtg
     2521 gtggttattg ggcaaatggt cgtcgtcctt tagcattagt tcgtacacat tcaaaaaaag
     2581 cattaatgcg ttatgaagat gtttacatgc ctgaagttta taaagccatt aatattgcac
     2641 agaatactgc atggaaaatc aacaagaaag ttttagcagt tgcaaatgtt attactaaat
     2701 ggaaacattg tcctgttgaa gatattcctg caattgaacg tgaagaatta ccaatgaaac
     2761 ctgaagatat tgatatgaat cctgaagcat taacagcatg gaaacgtgca gcagcagccg
     2821 tttatcgtaa agataaagca cgtaaatcac gtcgtatttc attagagttc atgttagagc
     2881 aagcaaacaa gttcgcaaat cataaagcca tttggtttcc ttataatatg gattggcgtg
     2941 gtcgtgttta tgcagtttca atgtttaatc ctcaaggtaa tgatatgacc aaaggtttat
     3001 taaccttagc taaaggtaaa cctattggta aagaaggtta ttattggtta aaaatccatg
     3061 gtgcaaattg tgcaggtgtt gataaagttc cttttccaga acgtattaag ttcattgaag
     3121 aaaatcatga aaatattatg gcatgtgcta aatcaccatt agaaaataca tggtgggcag
     3181 aacaagattc acctttttgt tttttagcct tttgttttga atatgcaggt gttcaacatc
     3241 atggtttatc atataattgt tcattaccat tagcatttga tggttcatgt tcaggtattc
     3301 agcatttttc agcaatgtta cgtgatgaag ttggtggtcg tgcagttaac ttattacctt
     3361 cagaaacagt tcaagatatc tatggtattg ttgctaaaaa agttaatgaa atcttacagg
     3421 cagatgccat taatggtaca gataatgaag ttgttacagt tacagatgaa aatacaggtg
     3481 aaatatcaga aaaagttaaa ttaggtacca aagcattagc aggtcaatgg ttagcatatg
     3541 gtgttacacg ttcagttact aagcgttcag ttatgacatt agcatatggt tcaaaagagt
     3601 tcggttttcg tcaacaggtt ttagaagata ccattcaacc tgcaattgat tcaggtaaag
     3661 gtttaatgtt tactcaacct aatcaagcag caggttatat ggccaaatta atatgggaat
     3721 cagtttcagt tacagttgtt gcagcagttg aagcaatgaa ttggttaaaa tcagcagcca
     3781 aattattagc agcagaagtt aaagataaaa aaacaggtga aatcttacgt aagcgttgtg
     3841 cagttcattg ggttacacct gatggttttc ctgtttggca agaatataaa aaacctattc
     3901 aaacacgttt aaacttaatg tttttaggtc agttccgttt acaacctact attaatacaa
     3961 acaaagattc agaaattgat gcacataaac aagaatcagg tattgcacct aacttcgttc
     4021 attcacaaga tggttcacat ttacgtaaaa cagttgtttg ggcacatgaa aaatatggta
     4081 ttgaatcatt tgcattaatt cacgattcat ttggtaccat tccagcagat gcagcaaact
     4141 tattcaaagc agttcgtgaa acaatggttg atacatatga atcatgtgat gttttagcag
     4201 acttctatga tcagttcgca gatcagttac atgaatcaca gttagataaa atgcctgcat
     4261 tacctgccaa aggtaactta aacttacgtg atattttaga atcagacttc gcatttgcct
     4321 aataaggcgc gccccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     4381 cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg
     4441 ggtgggcctt tctgcgttta tagctattac tcgaacccct agcccgctct tatcgggcgg
     4501 ctaggggttt tttgtatcga taccgtcgac ctcgaggggg ggcccggtac cagcttttgt
     4561 tccctttagt gagggttaat ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4621 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4681 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4741 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4801 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4861 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4921 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4981 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5041 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5101 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5161 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5221 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5281 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5341 tcgccactgg cagcagcaac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5401 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5461 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5521 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5581 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5641 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5701 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5761 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5821 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5881 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5941 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6001 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6061 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6121 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6181 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6241 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6301 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6361 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6421 ctcatcattg gaaaac
//
