LOCUS       sequence-120566-        8735 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Punc-4-QF-3'UTR-unc-54 (The endogenous SphI, KpnI and SacI sites in 
            QF are removed, and no SL2-mcherry).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8735)
  AUTHORS   Wei X, Potter CJ, Luo L, Shen K
  TITLE     Controlling gene expression with the Q repressible binary expression
            system in Caenorhabditis elegans.
  JOURNAL   Nat Methods. 2012 Mar 11;9(4):391-5. doi: 10.1038/nmeth.1929.
  PUBMED    22406855
REFERENCE   2  (bases 1 to 8735)
  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"; doi: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2012-03-11- 11"; volume: "9"; 
            issue: "4"; pages: "391-5"
COMMENT     Sequence Label: XW20
FEATURES             Location/Qualifiers
     source          1..8735
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(6254..6276)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6376..6395
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6477..6495)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6563..6667
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6668..6736,6737..7528)
                     /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(6886..6905)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7699..8287
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8188..8207
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8441..8458
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8575..8596
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8611..8616,8617..8634,8635..8641)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8649..8665
                     /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)."
     primer_bind     8654..8676
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgca gctctgagac catgttgatg ggaaattata
       61 tttgaaaact tttgaataaa gttgtataat tgtttttatt ctttaaccta tttatgaatg
      121 ctgtcccgga actgggatat aatttctaac ttgacacaat tattccacct ctgtcttcaa
      181 ggcgacctac acctacctcg cgtctaggtc caacagaaac actgttgaat gaaatcctcg
      241 aattgcaaca atataggatg ctcaggagac tataattcaa aaaacccaac ccaaattagc
      301 ataagcttcc ccaaattgga acagtgaaat actcaatcaa tcactctggc ggttttctcc
      361 acaaagaaca cgaaattgga aaaaggaaaa gattgatttg tttgaaggcg tgaggaaatc
      421 gaaaaaaaat tgtttcaaaa caaatgaaat attttaacga ataaacccag gaatgccact
      481 ttttggtgcc cgcctacaaa aaagtcaact gttcttaaac tcctacatag ttgccctctc
      541 taacgaatgt tgcttttttt aaattttcat tgaatttgat tatcgtctgt atgcgcctac
      601 tctatcagtg gaaaacatgc ccatttccca caaatttact cgtttggagc tcaaaatgtt
      661 aaatggggca tcagggaatt ggcatctcac agaaagtggc ggatgagtta tatgtaataa
      721 taatggtggc tgaagtggaa accgcaaaaa aaggatgtgt atttgagatt ttgcaacgga
      781 atttgaatca gatacacaaa atgagagaaa tttatagatt ctttaaaatc cacctgaagt
      841 cggcaaattt ttgaaaggtc tgttcgttca gtttttcata agttttatga attttgattt
      901 actactgaag ttcgagaaaa gccctttaac cataccaata agaaattatg ttaactttac
      961 aagaatatta acaaaaaatt attaaaaaac tgtgttcccc tggcattatg atatttccac
     1021 cgtcagttct ctattaaaac ttcaactcta cacagtttca actctcaaat gagctgtcca
     1081 tcttttgcct tcaggaaatt atgctaatct gctgatttcg cgcactttca ccacactatt
     1141 tccaaataga aggaggggtg gtgcaacaca cgagacactg caaaaaggtt cagtgaagat
     1201 ttatctactt gtgagatgag agcaggaggg gtgggagaag tgtgatgatt gacgggatga
     1261 caagaacagt gatgtggtgt cgaaatggaa gatgcgctaa tttgtggagt tttatgaaat
     1321 ttgaataatt acatttataa catctactag ttaggcgtct gtgtaggtat tgcctgcctg
     1381 cctacaaggc atatgtacga ggcaggttaa acctcctaga aagctacggt actttcaaac
     1441 cacataacat ttatactggt gtcttttcgc gtcttcttct tgaacatgta gaccctcaaa
     1501 aagctaagtt ttacactgtg tagtttgctt tgaaatctac aaaaattgga ggcacgcctt
     1561 ctggactttg ccgacacgcc tactttctac ctaccaactg cctacttcca cagtctagcg
     1621 tgaactccaa taccatttta aaaattgttt tatctcaaat tttcaatttt taaatccaat
     1681 ttcccagttc taaaaaaaaa attttttttt taatttaaaa actctcatcc tttttaatgc
     1741 tcgttcccat tcggcaagac gtgtaccaaa cacaaaacaa attgactggt gatccatctc
     1801 gaatttctcg tttcaaatta gtcgttgcta tatatattgt cttatgtgtg gaaaacgaat
     1861 aagaaaaatt agagaaagat gagagagcag gggcagaatg aaaatttgca tggaaattga
     1921 agaagatcac tgctttttcc gatgaatgat gactaaatag aatgagttga gaggaaaata
     1981 atcataattc taattttaat tagaaatatg atatggaaag taagcaaaac cacaaggctt
     2041 ttgctctgaa cttatttccg aaaaaaaaag tgaaactgaa agttagaatt ttcaaaataa
     2101 atattgcgag agtcaacctt cagaaaaatt ggcgagaaat caaatttgaa ttctctgaaa
     2161 atttagttaa aatacatatg ttatccttct ttgttttttg gccttagcct gccttcaagg
     2221 cgacctgagc ctatgcgggt gcctaatacc gaaacacaca taattttttt ttttaatctt
     2281 taaatactgt aacgagtcgc gagatccagc gagacccata gaatatgttg agcgctgcaa
     2341 acaatcgtat aactcttgtc caaatttttc cactctgtat ttttgcaagc agttcaacaa
     2401 atggtcatca tttttataac ccatatctat attcgaatct gacataacaa ataatactaa
     2461 gcacatcctc ccacatctct tttcattcaa gttgcgcaat tcccctaata actatggatg
     2521 gaggggggac cgagcccaaa gattattcaa aacgataacg actttttcat gcggccaacg
     2581 aaaaataaaa cttttctcat attctcctat ttcccatttc aagaattaca tccccatggg
     2641 acgttgaaat cacatcatat tagtatatgt taattgttct tttcactgga atatcgccac
     2701 gccccccttc cggtggctca ttggtttcga gaagaaggcg gagccgaagt gggttcttct
     2761 ttcttttttt tgtggtagaa gtctagcagg attaagctac tattccatta gaggatcttc
     2821 ttcttccaaa aagtgaaagg cgcgcctcta gaggatcccc gggattggcc aaaggaccca
     2881 aaggtatgtt tcgaatgata ctaacataac atagaacatt ttcaggagga cccttggcta
     2941 gccatgccac caaaacgcaa gacactcaat gccgctgccg aagccaatgc ccacgctgat
     3001 ggccatgctg atggcaatgc tgatggtcat gtcgctaaca ctgcagcaag cagcaacaac
     3061 gcccgttttg cggacttgac caacattgac acacccggcc tcggccctac cacgacgacg
     3121 ttacttgtcg agcccgctcg ttcgaaacgc cagagagtct cgagggcctg tgatcagtgt
     3181 cgagctgcac gtgaaaagtg tgatggaatc cagccggctt gcttcccctg tgtgtcgcag
     3241 ggccggtcgt gtacctacca ggccagtccc aagaagcgag gagtccagac gggctacatc
     3301 cgcactctcg aactggctct ggcttggatg ttcgagaacg ttgcccgcag cgaggacgcc
     3361 ctccacaatc ttttggtccg tgatgctggc cagggcagcg ctctcctggt cggcaaagac
     3421 tcgcctgctg cagaacgcct ccatgcaaga tgggcgacga gtcgagtcaa caaaagcatc
     3481 acccgtcttc tctcaggtca ggccgcacaa gatccatctg aagacggcca atccccgtcc
     3541 gaagacataa atgtccaaga tgccggggca aagacatccg acttccctca tgcgcctcac
     3601 ttgactttct cggcgcctaa gtctagtaca gctgagacac gcactctacc aggcccggtc
     3661 cgaccaccta tttcggcaaa caccctggaa aacaaccttc agccagatgg aaccgggata
     3721 gggaagctac cacccaatca ttggcgcctg ctggatatct acttttccta cacgcattct
     3781 tggctcccta tcctcgagaa gaaagacatg taccaagcat tgtaccagta ctctgaacaa
     3841 ggctcgttgc ttccctctgc gaatgtcgag tctggcgttc atgccgagct ttggagcgcg
     3901 ctcgccctgg cgtccttcca ggctgctgct actgctgcat cgagtgctac gggtccagct
     3961 tcagctgctc atggccatga caatgccatc aatccttcac ctgcagacat atctgacaca
     4021 gcccgaaagc tcataccttt ggaaagcggg ccgttccagg ttcagcactg cagagcgttg
     4081 ctgcttcttt gtctcgtaag ccttgggcgg gatgattggg agtctgcttg gttgctggtt
     4141 ggctttgcgg tccgcgtcct acttgttgtt cgcacccagt tgcctcctga tgatgaccgg
     4201 ccacgaccaa gaatgcgtgc gctgctcgtc gcgtgcttca tcgtggatac cattgtgtct
     4261 atgagacaca acgtgccggc ccatctcaag ccagacgaca ttgcggatct gccgttacct
     4321 gaagacggtc aagatcaatg ggagccgtgg acaccatgtg agggcttagg cggtgaacac
     4381 accatgctgc aaatgttgag gaacccggca taccctttaa gcacattcaa ccacctatat
     4441 ggcgtgacca agctggttgc tttggagctt ctgccaagaa tacgaacatc ttcacagaac
     4501 gctcccttgg agttcaggtc gcggttgcag caggtaatcg gccacaattc tcccttcagc
     4561 gtctttgtcc tttcccagga tacagcatcg gcttttgtgc ctactgcata ccttacccgt
     4621 accgtttatt tatgggcagc tgccttttct gagcctctca acgaacacta ctcgcatctt
     4681 ctgatcgaaa ctcttgatca gtatcagaag cggtttggta catatgcaat cccacctctg
     4741 atcccttctc ttctagactc ccttcttgct ttaaagaaac aatcacattc ttcagagcgg
     4801 catcgaaggc acttggaaga gcttttcccc gcctactcct ccatttggcc tcggggaggc
     4861 cgacacagca atactggcct ccaacccata cgacaacttg agcttccgcc gactgcgact
     4921 gccactgcaa gtatcatgcc ccatgtcatg gaacagcccc tgtcaacgtc aataaatccg
     4981 gtcaatgatc ggtttaatgg aataccgaat cctaccccct acaatagcga tgcggcccta
     5041 gacgcaatta ctcaaaccaa tgactacgga tcagtcaata cccatggcat tctcagtaca
     5101 tatccaccac ctgccaccca cttgaacgag gcctcggtgg cccttgcgcc tggaggtgct
     5161 cctcccagac cacccccgcc atacgttgat agtacaacga accatcctcc ttaccatagt
     5221 aacctcgtcc ccatggccaa tttcggatat tcgactgtag actatgatgc gatggtggat
     5281 gatctggcat cgatcgagta cacggatgcg gttgacgttg atccgcagtt catgacgaac
     5341 ctcgggtttg tcccagggtg taattttagc gatatcaaca cgtatgagca ataggctagc
     5401 gtcgacggta ccatggtatt gatatctgag ctccgcatcg gccgctgtca tcagatcgcc
     5461 atctcgcgcc cgtgcctctg acttctaagt ccaattactc ttcaacatcc ctacatgctc
     5521 tttctccctg tgctcccacc ccctattttt gttattatca aaaaaacttc ttcttaattt
     5581 ctttgttttt tagcttcttt taagtcacct ctaacaatga aattgtgtag attcaaaaat
     5641 agaattaatt cgtaataaaa agtcgaaaaa aattgtgctc cctcccccca ttaataataa
     5701 ttctatccca aaatctacac aatgttctgt gtacacttct tatgtttttt ttacttctga
     5761 taaatttttt ttgaaacatc atagaaaaaa ccgcacacaa aataccttat catatgttac
     5821 gtttcagttt atgaccgcaa tttttatttc ttcgcacgtc tgggcctctc atgacgtcaa
     5881 atcatgctca tcgtgaaaaa gttttggagt atttttggaa tttttcaatc aagtgaaagt
     5941 ttatgaaatt aattttcctg cttttgcttt ttgggggttt cccctattgt ttgtcaagag
     6001 tttcgaggac ggcgtttttc ttgctaaaat cacaagtatt gatgagcacg atgcaagaaa
     6061 gatcggaaga aggtttgggt ttgaggctca gtggaaggtg agtagaagtt gataatttga
     6121 aagtggagta gtgtctatgg ggtttttgcc ttaaatgaca gaatacattc ccaatatacc
     6181 aaacataact gtttcctact agtcggccgt acgggccctt tcgtctcgcg cgtttcggtg
     6241 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     6301 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     6361 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     6421 aaataccgca cagatgcgta aggagaaaat accgcatcag gcggccttaa ttaagggcct
     6481 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     6541 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6601 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6661 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6721 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6781 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6841 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6901 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6961 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     7021 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     7081 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     7141 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7201 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     7261 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     7321 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     7381 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     7441 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     7501 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     7561 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7621 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7681 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7741 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7801 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     7861 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7921 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7981 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     8041 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag
     8101 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     8161 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     8221 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     8281 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     8341 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     8401 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     8461 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     8521 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     8581 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     8641 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctgggggt gggggggggg
     8701 ttgggggtgg ggtgttgttg gggtgtgttt tgtgt
//
