LOCUS       sequence-64991-e        8374 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8374)
  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 8374)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-9-QFMD184-475
FEATURES             Location/Qualifiers
     source          1..8374
                     /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"
     primer_bind     complement(7980..7997)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7980..7996)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7989..8011)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(8205..8224)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8324..8346
                     /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 aggtttgcgt cagttggagc
     6241 tccctcctac cgccacggct acggcctcga taatgccgca cgtgatggag cagcctctca
     6301 gtaccagcat taaccccgtc aacgaccgct tcaacggtat tcccaacccc actccgtata
     6361 actccgatgc agctctcgat gctatcactc agaccaacga ttatggaagc gtaaatacac
     6421 atggtatcct ctctacttac ccgccaccgg ctacgcacct taatgaagct tccgtcgctc
     6481 tcgctcccgg tggcgccccc ccccgaccgc ctcctccgta tgttgacagc acgaccaatc
     6541 acccgccgta ccactcgaat ctggttccaa tggcgaactt tggttactcg accgttgatt
     6601 acgatgccat ggttgacgat ttggctagca ttgaatacac ggacgctgtg gatgtcgacc
     6661 cacagtttat gaccaatctg ggattcgttc ctggatgtaa cttctccgac attaatacat
     6721 acgaacagtg aggccgcgtt aaccgatccg tcgaccatga agatcaagat cattgccccg
     6781 ccagagcgca agtactctgt ctggatcggt ggctccatcc tggcttcgct gtccaccttc
     6841 cagcagatgt ggatctccaa gcaggagtac gacgagtccg gcccctccat tgtgcaccgc
     6901 aagtgcttct aagaaggatc gcttgtctgg gcaagaggat caggatcggg atggtcttga
     6961 ttctgctgga ggaggaggag gagaagtcga ggaagcagca gcgaaagtgc aagtgcgagt
     7021 ggtggaagtt tggagtgcag cacaacaaaa tcaacaacaa caccaactac aagatgaaaa
     7081 gagcggaacc acctgcacac catcatcact atcatcatcg ttttgggcgc atgttgtgtg
     7141 gttccagcgt attaatataa ttaatttatt ccacatgaga tatgatatga tatactatgt
     7201 attttttgtt ttttttttat ttgtaaacct ttaatataac aagaactaca aaaaatgaaa
     7261 atgagcgaaa atgcatattc tgccattcca cacacacacc aacaacaccc aacacacgca
     7321 cacccacaag cttacacaca cacattcgcg gcatgacaag gacatcaaga taaagaagaa
     7381 cttaaagaag atatttccca aagcgcaaaa agaacacaca cacattgcaa aacacaaaca
     7441 acacactagc gttttgtaca attcgtcagc aaccttatgt attatttttt aattatgatg
     7501 taattataaa caaagtgaaa caaaaatatg aaaacaaaaa ggaaaatcaa atctgtcttc
     7561 tctttctccc gctctcctcg ctctctgctg ctaacctcgc cctctcctct ctcatctttt
     7621 tgtctgtctc tcttccacat ttttgccggc cggcaaaata ataacccaca cacactcaca
     7681 cttggctgca gtttcgcgtg cgatattcac acacattcaa gcatacaagc atacatacat
     7741 atgtattttt tttttatttg tacacttttc taattgcatg cgtatcgatt gataagttta
     7801 cgcctgaaaa tgttaattaa aatgtgaaaa tgcaactgaa aaactgatga aatgaaacaa
     7861 caacaagcga acaatttgct acatgtgtat tgtctaacaa ccgttactgc aacggttgct
     7921 tcggaaaagg gtgaagagga agagggcacg aggtcgacct gcaggcatgc aagcttggca
     7981 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     8041 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     8101 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt
     8161 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
     8221 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     8281 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     8341 cagaggtttt caccgtcatc accgaaacgc gcga
//
