LOCUS       sequence-66365-e        9449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9449)
  AUTHORS   Riabinina O, Luginbuhl D, Marr E, Liu S, Wu MN, Luo L, Potter CJ
  TITLE     Improved and expanded Q-system reagents for genetic manipulations.
  JOURNAL   Nat Methods. 2015 Jan 12. doi: 10.1038/nmeth.3250.
  PUBMED    25581800
REFERENCE   2  (bases 1 to 9449)
  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 
            Methods. 2015 Jan 12. doi: 10.1038/nmeth.3250."
COMMENT     Sequence Label: pAC-15-LexABD-G4MD-QFAD
FEATURES             Location/Qualifiers
     source          1..9449
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2277..4790
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             4815..4835
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4845..5096,5097..5450)
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5'-CTGTATATATATACAG-3'."
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     primer_bind     complement(9055..9072)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9055..9071)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9064..9086)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(9280..9299)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9399..9421
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgaattctat attctaaaaa cacaaatgat acttctaaaa aaaaatcatg
     2281 aatggcatca actctgaatc aaatctttgc agatgcacct acttctcatt tccactgtca
     2341 catcattttt ccagatctcg ctgcctgtta tgtggcccac aaaccaagac acgttttatg
     2401 gccattaaag ctggctgatc gtcgccaaac accaaataca taatgaatat gtacacattc
     2461 gagaaagaag cgatcaaaga agcgtcttcg ggcggagtag gagaatgcgg aggagaagga
     2521 gaacgagctg atctagtatc tctccacaat ccaatgccaa ctgaccaact ggccatattc
     2581 ggagcaattt gaagccaatt tccatcgcct ggcgatcgct ccattcttgg ctatatgttt
     2641 ttcaccgtta cccggggcca ttttcaaaga ctcgtcggca agataagatt gtgtcactcg
     2701 ctgtctctct tcatttgtcg aagaatgctg aggaatttcg cgatgacgtc ggcgagtatt
     2761 ttgaagaatg agaataattt gtatttatac gaaaatcagt tagtggaatt ttctacaaaa
     2821 acatgttatc tatagataat tttgttgcaa aatatgttga ctatgacaaa gattgtatgt
     2881 atataccttt aatgtattct cattttctta tgtatttata atggcaatga tgatactgat
     2941 gatattttaa gatgatgcca gaccaaaagg cttgaatttc tgcgtctttt gccgaacgca
     3001 gtgcatgtgc aattgttgtt ttttggaata ttcaattttc ggactgtccg ctttgatttc
     3061 agtttcttgg cttattcaaa aagcaaagta aagccaaaaa agcgagatgg caataccaaa
     3121 tgcggcaaaa cggtagtgga aggaaagggg tgcggggcag cggaaggaag ggtggggcgg
     3181 ggcgtggcgg ggtctgtggc tgggcgcgac gtcaccgacg ttggagccac tcctttgacc
     3241 atgtgtgcgt gtgtgtatta ttcgtgtctc gccactcgcc ggttgttttt ttctttttat
     3301 gctgcgctct ctctagcgcc atctcgctta cgcatgctca acgcaccgca tgttgccgtt
     3361 tccttttatg cgtcattttg gctcgaaata ggcaattatt taaacaaaga ttagtcaacg
     3421 aaaacgctaa aataaataag tctacaatat ggttacttat tgccatgtgt gtgcagccaa
     3481 cgatagcaac aaaagcaaca acacaggtgg ctttccctct ttcacttttt gtttgcaagc
     3541 cgcgtgcgag caagacggca cgaccggcaa acgcaattac gctgacaaag agcagacgaa
     3601 gttttggcga aaaacatcaa ggcgcctgat acgaatgcat ttgcaataac aattgcgata
     3661 tttaatattg tttatgaagc tgtttgactt caaaacacac aaaaaaaaaa ataaaacaaa
     3721 ttatttgaaa gagaattagg aatcggacgc ttatcgttag ggtaacaaca agaaatgctt
     3781 actgagtcac agcctctgga aaactgccgc aagccagaga gagagagaaa aagagggaga
     3841 gcagcttaga ccgcatgtgc ttgtgtgtga ggcgtctctc tcttcgtctc tgttgcgcaa
     3901 acgcatagac tgcactgaaa aaatcgatta cctatttttt atgaatgaat atttgcacta
     3961 ttactattca aaactattaa gatagcaatc acattcaata gccaaatact ataccacctg
     4021 agcgatgcaa cgaaatgatc aatttgagca aaaatgctgc atatttagga cggcatcatt
     4081 atagaaatgc ttcttgctgt gtacttttct ctcgtctggc agctgtttcg ccgttattgt
     4141 taaaaccggc ttaagttagg tgtgttttct acgactagct agtgaatgcc ctactagaag
     4201 atgtgtgttg cacaaaatgt ccctggaata accaatttga agtgcagata gcagtaaacg
     4261 taagctaata tgaatattat ttaactgtaa tgttttaata tcgctggaca ttactaataa
     4321 acccactata aacacatgta catatgtatg ttttggcata caatgagtag ttggggaaaa
     4381 aatgtgtaaa agcaccgtga ccatcacagc ataaagataa ccagctgaag tatcgaatat
     4441 gagtaacccc caaattgaat cacatgccgc aactgatagg acccatggaa gtacactctt
     4501 catggcgata tacaagacac acacaagcac gaacacccag ttgcggagga aattctccgt
     4561 aaatgaaaac ccaatcggcg aacaattcat acccatatat ggtaaaagtt ttgaacgcga
     4621 cttgagagcg gagagcattg cggctgataa ggttttagcg ctaagcgggc tttataaaac
     4681 gggctgcggg accagttttc atatcactac cgtttgagtt cttgtgctgt gtggatactc
     4741 ctcccgacac aaagccgctc catcagccag cagtcgtcta atccagagac cccggatcga
     4801 attccaaaat gccacccaag aagaagcgaa aagtagaaga tccaatgaag gctctcacgg
     4861 cccgacaaca ggaagttttt gatttgatac gggatcatat atcccaaacg ggtatgcctc
     4921 cgacccgcgc agagatagca cagcgactgg gctttcgatc gcctaacgcc gcggaggagc
     4981 acttgaaggc actggcccgc aagggtgtca ttgaaatcgt gtccggtgcg agccgcggaa
     5041 tccggctgtt gcaggaggag gaagagggcc tgccactggt gggacgcgtg gccgctggcg
     5101 agccgctgct ggcccagcaa cacatagagg gacactatca ggtggacccc tccttgttta
     5161 agccaaatgc tgatttcctg ttgcgggtgt cgggaatgtc catgaaggac atcggtatta
     5221 tggatggtga cctcctcgcc gtccataaga cacaggatgt caggaacggc caggtagtcg
     5281 ttgccaggat agacgatgag gtcactgtga aacgtctcaa gaagcaaggc aataaggtcg
     5341 agctgctgcc ggagaatagc gagttcaagc cgatcgtggt ggatctgcga cagcagtcct
     5401 ttactatcga gggcttggcc gtgggtgtga tccgcaacgg agattggctg ggatccatcg
     5461 acagtgcggc ccaccacgac aacagcacca ttccactgga cttcatgcct cgtgacgcac
     5521 tgcatggatt tgactggagc gaggaggatg atatgagcga cggactccca ttcctgaaga
     5581 ccgaccccaa taacaatggt ttctttggcg acggcagcct gctgtgtatc ctgcgaagta
     5641 ttggttttaa gccggaaaat tacacgaaca gtaatgtcaa tcgtcttccc acaatgatca
     5701 ccgaccgtta cacgctggcc agtcgcagta ctacaagccg cctgctgcaa agctatctta
     5761 acaacttcca cccctactgc cccatagtgc atagtcctac tctcatgatg ctttacaaca
     5821 atcagattga gatagcatcg aaggaccagt ggcagatcct cttcaactgc attctcgcca
     5881 tcggcgcttg gtgtattgaa ggcgagagca cagatatcga cgtgttttac tatcaaaacg
     5941 ccaagtcgca cctcacctcg aaggtcttcg aaagtggaag cattatcctc gtgacagccc
     6001 tccatctgct gagtcgttat acccagtggc gtcagaaaac taatacatcc tacaactttc
     6061 attcgttcag cattcggatg gcgattagcc tgggacttaa tcgtgacctc ccaagtagct
     6121 tcagcgattc gtcgatcctg gaacagcgcc ggaggatttg gtggagtgtg tacagctggg
     6181 agatccagct gtccctcttg tacggacgca gcattcagct gagtcagaac acgatttcct
     6241 tcccgtccag cgtggacgac gttcaacgga caactacggg cccgaccatc taccacggta
     6301 tcattgagac ggcccgcttg cttcaagtgt ttacgaagat ttatgagctg gataagacag
     6361 tgaccgcgga gaagagtccg atctgcgcca agaaatgcct gatgatctgc aatgagatag
     6421 aggaggtgag ccgccaggca cccaagttcc tgcagatgga cataagcact acggcgctga
     6481 ctaatctcct caaagaacac ccctggctga gtttcacccg cttcgagctg aagtggaaac
     6541 agctgagtct gattatatac gttttgcggg actttttcac aaattttacg cagaagaagt
     6601 cgcagcttga gcaagatcag aacgaccatc agtcctacga ggtaaagagg tgcagcataa
     6661 tgctctccga cgcagcccag cgcaccgtga tgtcggtgtc gtcctatatg gacaaccata
     6721 atgtgacccc ttatttcgcc tggaattgct cctactattt gttcaacgcc gtgttggtcc
     6781 caatcaagac tcttctgagc aactccaaat cgaacgccga gaataacgag acagcacagc
     6841 tcttgcagca aattaacacc gtgttgatgc tgttgaagaa actcgctaca ttcaagattc
     6901 agacttgcga aaagtacatc caggtgttgg aagaggtgtg cgccccattc cttctttcgc
     6961 aatgcgcaat tcccctcccg cacataagtt acaataattc caacggctcc gctatcaaga
     7021 atatcgtcgg ctccgctaca atcgcgcagt atcccacgct gcctgaggaa aatgttaaca
     7081 acatatcggt gaagtatgta agtccaggca gcgtcggccc gagccctgtg ccgctgaaaa
     7141 gcggagcctc cttcagcgat ctggtcaagc tgctcagcaa ccgcccacca tcccgcaact
     7201 cccctgtcac tataccccga tcgaccccgt cgcatcgtag cgttacccca ttcctgggac
     7261 aacagcagca actgcagagt ctggtgccac ttacgcgtca gttggagctc cctcctaccg
     7321 ccacggctac ggcctcgata atgccgcacg tgatggagca gcctctcagt accagcatta
     7381 accccgtcaa cgaccgcttc aacggtattc ccaaccccac tccgtataac tccgatgcag
     7441 ctctcgatgc tatcactcag accaacgatt atggaagcgt aaatacacat ggtatcctct
     7501 ctacttaccc gccaccggct acgcacctta atgaagcttc cgtcgctctc gctcccggtg
     7561 gcgccccccc ccgaccgcct cctccgtatg ttgacagcac gaccaatcac ccgccgtacc
     7621 actcgaatct ggttccaatg gcgaactttg gttactcgac cgttgattac gatgccatgg
     7681 ttgacgattt ggctagcatt gaatacacgg acgctgtgga tgtcgaccca cagtttatga
     7741 ccaatctggg attcgttcct ggatgtaact tctccgacat taatacatac gaacagtgat
     7801 gacgtcggcc gcgttaaccg atccgtcgac catgaagatc aagatcattg ccccgccaga
     7861 gcgcaagtac tctgtctgga tcggtggctc catcctggct tcgctgtcca ccttccagca
     7921 gatgtggatc tccaagcagg agtacgacga gtccggcccc tccattgtgc accgcaagtg
     7981 cttctaagaa ggatcgcttg tctgggcaag aggatcagga tcgggatggt cttgattctg
     8041 ctggaggagg aggaggagaa gtcgaggaag cagcagcgaa agtgcaagtg cgagtggtgg
     8101 aagtttggag tgcagcacaa caaaatcaac aacaacacca actacaagat gaaaagagcg
     8161 gaaccacctg cacaccatca tcactatcat catcgttttg ggcgcatgtt gtgtggttcc
     8221 agcgtattaa tataattaat ttattccaca tgagatatga tatgatatac tatgtatttt
     8281 ttgttttttt tttatttgta aacctttaat ataacaagaa ctacaaaaaa tgaaaatgag
     8341 cgaaaatgca tattctgcca ttccacacac acaccaacaa cacccaacac acgcacaccc
     8401 acaagcttac acacacacat tcgcggcatg acaaggacat caagataaag aagaacttaa
     8461 agaagatatt tcccaaagcg caaaaagaac acacacacat tgcaaaacac aaacaacaca
     8521 ctagcgtttt gtacaattcg tcagcaacct tatgtattat tttttaatta tgatgtaatt
     8581 ataaacaaag tgaaacaaaa atatgaaaac aaaaaggaaa atcaaatctg tcttctcttt
     8641 ctcccgctct cctcgctctc tgctgctaac ctcgccctct cctctctcat ctttttgtct
     8701 gtctctcttc cacatttttg ccggccggca aaataataac ccacacacac tcacacttgg
     8761 ctgcagtttc gcgtgcgata ttcacacaca ttcaagcata caagcataca tacatatgta
     8821 tttttttttt atttgtacac ttttctaatt gcatgcgtat cgattgataa gtttacgcct
     8881 gaaaatgtta attaaaatgt gaaaatgcaa ctgaaaaact gatgaaatga aacaacaaca
     8941 agcgaacaat ttgctacatg tgtattgtct aacaaccgtt actgcaacgg ttgcttcgga
     9001 aaagggtgaa gaggaagagg gcacgaggtc gacctgcagg catgcaagct tggcactggc
     9061 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     9121 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     9181 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca
     9241 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     9301 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     9361 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     9421 gttttcaccg tcatcaccga aacgcgcga
//
