LOCUS       sequence-36641-e        6723 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 6723)
  AUTHORS   Stowers RS
  TITLE     An efficient method for recombineering GAL4 and QF drivers.
  JOURNAL   Fly (Austin). 2011 Oct 1;5(4).
  PUBMED    21857163
REFERENCE   2  (bases 1 to 6723)
  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: "Fly 
            (Austin). 2011 Oct 1;5(4)."
COMMENT     Sequence Label: pQFKan
FEATURES             Location/Qualifiers
     source          1..6723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(129..197,198..989)
                     /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(347..366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1902..1919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2072..2077,2078..2095,2096..2102)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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     2115..2137
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2134..2150
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2169..2189
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    complement(4947..4980)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(5083..5102)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5094..5903
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5161..5180)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     protein_bind    complement(5978..6011)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(6078..6097)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6079..6097)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6107..6124)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6107..6123)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6116..6138)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(6264..6720)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6402..6423)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6614..6633
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggccactgtt taaacttggc
     2221 gcgcccaaca tgccgcctaa acgcaagaca ctcaatgccg ctgccgaagc caatgcccac
     2281 gctgatggcc atgctgatgg caatgctgat ggtcatgtcg ctaacactgc agcaagcagc
     2341 aacaacgccc gttttgcgga cttgaccaac attgacacac ccggcctcgg ccctaccacg
     2401 acgacgttac ttgtcgagcc cgctcgttcg aaacgccaga gagtctcgag ggcctgtgat
     2461 cagtgtcgag ctgcacgtga aaagtgtgat ggaatccagc cggcttgctt cccctgtgtg
     2521 tcgcagggcc ggtcgtgtac ctaccaggcc agtcccaaga agcgaggagt ccagacgggc
     2581 tacatccgca ctctcgaact ggctctggct tggatgttcg agaacgttgc ccgcagcgag
     2641 gacgccctcc acaatctttt ggtccgtgat gctggccagg gcagcgctct cctggtcggc
     2701 aaagactcgc ctgctgcaga acgcctgcat gcaagatggg cgacgagtcg agtcaacaaa
     2761 agcatcaccc gtcttctctc aggtcaggcc gcacaagatc catctgaaga cggccaatcc
     2821 ccgtccgaag acataaatgt ccaagatgcc ggggcaaaga catccgactt ccctcatgcg
     2881 cctcacttga ctttctcggc gcctaagtct agtacagctg agacacgcac tctaccaggc
     2941 ccggtccgac cacctatttc ggcaaacacc ctggaaaaca accttcagcc agatggtacc
     3001 gggataggga agctaccacc caatcattgg cgcctgctgg atatctactt ttcctacacg
     3061 cattcttggc tccctatcct cgagaagaaa gacatgtacc aagcattgta ccagtactct
     3121 gaacaaggct cgttgcttcc ctctgcgaat gtcgagtctg gcgttcatgc cgagctctgg
     3181 agcgcgctcg ccctggcgtc cttccaggct gctgctactg ctgcatcgag tgctacgggt
     3241 ccagcttcag ctgctcatgg ccatgacaat gccatcaatc cttcacctgc agacatatct
     3301 gacacagccc gaaagctcat acctttggaa agcgggccgt tccaggttca gcactgcaga
     3361 gcgttgctgc ttctttgtct cgtaagcctt gggcgggatg attgggagtc tgcttggttg
     3421 ctggttggct ttgcggtccg cgtcctactt gttgttcgca cccagttgcc tcctgatgat
     3481 gaccggccac gaccaagaat gcgtgcgctg ctcgtcgcgt gcttcatcgt ggataccatt
     3541 gtgtctatga gacacaacgt gccggcccat ctcaagccag acgacattgc ggatctgccg
     3601 ttacctgaag acggtcaaga tcaatgggag ccgtggacac catgtgaggg cttaggcggt
     3661 gaacacacca tgctgcaaat gttgaggaac ccggcatacc ctttaagcac attcaaccac
     3721 ctatatggcg tgaccaagct ggttgctttg gagcttctgc caagaatacg aacatcttca
     3781 cagaacgctc ccttggagtt caggtcgcgg ttgcagcagg taatcggcca caattctccc
     3841 ttcagcgtct ttgtcctttc ccaggataca gcatcggctt ttgtgcctac tgcatacctt
     3901 acccgtaccg tttatttatg ggcagctgcc ttttctgagc ctctcaacga acactactcg
     3961 catcttctga tcgaaactct tgatcagtat cagaagcggt ttggtacata tgcaatccca
     4021 cctctgatcc cttctcttct agactccctt cttgctttaa agaaacaatc acattcttca
     4081 gagcggcatc gaaggcactt ggaagagctt ttccccgcct actcctccat ttggcctcgg
     4141 ggaggccgac acagcaatac tggcctccaa cccatacgac aacttgagct tccgccgact
     4201 gcgactgcca ctgcaagtat catgccccat gtcatggaac agcccctgtc aacgtcaata
     4261 aatccggtca atgatcggtt taatggaata ccgaatccta ccccctacaa tagcgatgcg
     4321 gccctagacg caattactca aaccaatgac tacggatcag tcaataccca tggcattctc
     4381 agtacatatc caccacctgc cacccacttg aacgaggcct cggtggccct tgcgcctgga
     4441 ggtgctcctc ccagaccacc cccgccatac gttgatagta caacgaacca tcctccttac
     4501 catagtaacc tcgtccccat ggccaatttc ggatattcga ctgtagacta tgatgcgatg
     4561 gtggatgatc tggcatcgat cgagtacacg gatgcggttg acgttgatcc gcagttcatg
     4621 acgaacctcg ggtttgtccc agggtgtaat tttagcgata tcaacacgta tgagcaatag
     4681 gctagcgtcg actaaagcca aatagaaaat tattcagttc ctggcttaag tttttaaaag
     4741 tgatattatt tatttggttg taaccaacca aaagaatgta aataactaat acataattat
     4801 gttagtttta agttagcaac aaattgattt tagctatatt agctacttgg ttaataaata
     4861 gaatatattt atttaaagat aattgcgttt ttattgtcag ggagtgagtt tgcttaaaaa
     4921 ctcgtttaga tcggatccgc ggccgcataa cttcgtatag catacattat acgaagttat
     4981 ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     5041 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     5101 atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg
     5161 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg
     5221 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     5281 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     5341 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa
     5401 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     5461 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     5521 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     5581 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac
     5641 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     5701 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     5761 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     5821 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     5881 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca
     5941 ttacgctgac ttgacgggac ggcgcaagct catgaccata acttcgtata gcatacatta
     6001 tacgaagtta tgcagtcatg cgtctgagaa ttccacttaa ttaagaaggc cggcccttcc
     6061 tcagcgtggc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     6121 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     6181 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     6241 tgcgcagcct gaatggcgaa tggaaattgt aagcgttaat attttgttaa aattcgcgtt
     6301 aaatttttgt taaatcagct cattttttta accaataggc cgaaatcggc aaaatccctt
     6361 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     6421 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     6481 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     6541 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     6601 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     6661 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     6721 cag
//
