LOCUS       sequence-64985-e        8767 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 8767)
  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 8767)
  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-4-QFBDMD-G4AD
FEATURES             Location/Qualifiers
     source          1..8767
                     /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             6780..7124
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     primer_bind     complement(8373..8390)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8373..8389)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8382..8404)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(8598..8617)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8717..8739
                     /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 cccggatcca
     4801 acatgccacc caagcgcaaa acgcttaacg ctgcggctga ggctaacgct catgccgacg
     4861 gacacgccga cggaaacgcc gacggacacg tggccaatac ggccgcgtcc tcgaataatg
     4921 cgaggttcgc tgatctcact aacatcgata ctccgggtct gggacccaca actacgaccc
     4981 tgctcgtgga accagcacgc tcaaagcgtc aacgagtgtc ccgcgcatgc gaccagtgcc
     5041 gtgcagcccg agagaaatgc gacggaatac agcctgcgtg tttcccgtgc gtttcccagg
     5101 gaaggtcctg cacttatcag gcttcgccga aaaagagggg agttcaaacc ggttatattc
     5161 gtacgctgga gctcgccctc gcctggatgt ttgaaaatgt cgcgcgttcc gaagatgcct
     5221 tgcataacct cctcgtccgt gacgccggac aaggatcagc tctgctcgtt ggtaaagatt
     5281 cgccggctgc cgagcgactc catgcccgtt gggctactag ccgtgtcaat aagagcatta
     5341 cccgcctcct ctcgggccaa gccgctcaag acccttcgga agatggacag tcgccttcgg
     5401 aggatatcaa tgttcaagat gcgggagcaa aaacctcgga tttcccgcat gctccccacc
     5461 tcacgttcag cgcacctaaa agttcgactg cggagactag gactctccct ggtccagttc
     5521 gtcctccaat ctcggcgaac acactcgaaa ataatctgca gccggatggc actggaatag
     5581 gaaagctgcc tcctaaccac tggcgtttgt tggacattta tttctcgtat acccattcgt
     5641 ggcttccaat tttggaaaaa aaggatatgt atcaggctct ctaccaatac agcgagcagg
     5701 gaagcctcct tccgtcggcg aacgttgaaa gcggtgtaca tgctgaactg tggtcagcgt
     5761 tggcgcttgc gtcgtttcaa gccgctgcga ccgccgcgtc cagcgcgacg ggacctgcaa
     5821 gcgcagctca cggacacgat aatgcgatca acccgagtcc cgcggatatt tcggatacgg
     5881 caaggaaact catccctctt gaatcgggac catttcaagt ccagcattgc agggctctct
     5941 tgttgttgtg cctggttagc ctcggacgcg acgactggga atcggcctgg ctgcttgttg
     6001 gtttcgccgt ccgtgtcttg ttggtggttc gtacacagct gcccccagac gatgatcgtc
     6061 caaggccccg gatgagggct ttgctcgtgg cctgttttat tgttgacacg atcgtctcca
     6121 tgcgccataa tgtcccggct cacctgaaac ccgatgatat tgctgatttg ccgctgccag
     6181 aggatggcca ggatcagtgg gaaccttgga ccccctgcga aggtttggga ggagagcaca
     6241 caatgttgca gatgcttcgt aatccggctt atccactcag caccttcaat cacttgtacg
     6301 gtgtcactaa gttggtagct ctggagttgc tccctcgcat ccgtacctcg tcgcagaatg
     6361 cccctctgga atttagatcc cgactgcagc aagtcattgg tcataacagt ccattctcgg
     6421 tcttcgtcct gtcgcaagac acggcaagcg ctttcgtccc gacggcttat ctgactcgta
     6481 ctgtttacct ctgggcggcg gcgttcagcg aacccctgaa tgagcattat tcccacctct
     6541 tgattgagac gctcgaccag taccaaaaac gcttcggaac atatgccatt ccgcccctta
     6601 ttccgtcgct cctcgactcg ctcctggctc tcaaaaagca gagccattcg tcggagcgtc
     6661 acaggcgcca cctggaggag ctatttccgg cttattcgtc tatctggccg cgaggtggaa
     6721 ggcattcaaa cactggtctg cagccaattc cactgacccc gtctgctttg tttggtggcg
     6781 ccaattttaa tcaaagtggg aatattgctg atagctcatt gtccttcact ttcactaaca
     6841 gtagcaacgg tccgaacctc ataacaactc aaacaaattc tcaagcgctt tcacaaccaa
     6901 ttgcctcctc taacgttcat gataacttca tgaataatga aatcacggct agtaaaattg
     6961 atgatggtaa taattcaaaa ccactgtcac ctggttggac ggaccaaact gcgtataacg
     7021 cgtttggaat cactacaggg atgtttaata ccactacaat ggatgatgta tataactatc
     7081 tattcgatga tgaagatacc ccaccaaacc caaaaaaaga gtaaggccgc gttaaccgat
     7141 ccgtcgacca tgaagatcaa gatcattgcc ccgccagagc gcaagtactc tgtctggatc
     7201 ggtggctcca tcctggcttc gctgtccacc ttccagcaga tgtggatctc caagcaggag
     7261 tacgacgagt ccggcccctc cattgtgcac cgcaagtgct tctaagaagg atcgcttgtc
     7321 tgggcaagag gatcaggatc gggatggtct tgattctgct ggaggaggag gaggagaagt
     7381 cgaggaagca gcagcgaaag tgcaagtgcg agtggtggaa gtttggagtg cagcacaaca
     7441 aaatcaacaa caacaccaac tacaagatga aaagagcgga accacctgca caccatcatc
     7501 actatcatca tcgttttggg cgcatgttgt gtggttccag cgtattaata taattaattt
     7561 attccacatg agatatgata tgatatacta tgtatttttt gttttttttt tatttgtaaa
     7621 cctttaatat aacaagaact acaaaaaatg aaaatgagcg aaaatgcata ttctgccatt
     7681 ccacacacac accaacaaca cccaacacac gcacacccac aagcttacac acacacattc
     7741 gcggcatgac aaggacatca agataaagaa gaacttaaag aagatatttc ccaaagcgca
     7801 aaaagaacac acacacattg caaaacacaa acaacacact agcgttttgt acaattcgtc
     7861 agcaacctta tgtattattt tttaattatg atgtaattat aaacaaagtg aaacaaaaat
     7921 atgaaaacaa aaaggaaaat caaatctgtc ttctctttct cccgctctcc tcgctctctg
     7981 ctgctaacct cgccctctcc tctctcatct ttttgtctgt ctctcttcca catttttgcc
     8041 ggccggcaaa ataataaccc acacacactc acacttggct gcagtttcgc gtgcgatatt
     8101 cacacacatt caagcataca agcatacata catatgtatt ttttttttat ttgtacactt
     8161 ttctaattgc atgcgtatcg attgataagt ttacgcctga aaatgttaat taaaatgtga
     8221 aaatgcaact gaaaaactga tgaaatgaaa caacaacaag cgaacaattt gctacatgtg
     8281 tattgtctaa caaccgttac tgcaacggtt gcttcggaaa agggtgaaga ggaagagggc
     8341 acgaggtcga cctgcaggca tgcaagcttg gcactggccg tcgttttaca acgtcgtgac
     8401 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     8461 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     8521 ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     8581 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     8641 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     8701 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     8761 cgcgcga
//
