LOCUS       sequence-325425-        9582 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of 1 LoxN site in the genome of P. berghei.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9582)
  TITLE     Silvie lab unpublished
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9582)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pDownstream1Lox
FEATURES             Location/Qualifiers
     source          1..9582
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1732..2442
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             1732..2442
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1759..1787)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2382..2402
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2452..2505
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     protein_bind    complement(3733..3766)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     primer_bind     complement(6428..6448)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6428..6446)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6467..6483)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6467..6483)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6480..6502)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6491..6507
                     /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)."
     promoter        complement(join(6515..6521,6522..6539,6540..6545))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6560..6581
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6698..6715)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6869..7457)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6949..6968)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7628..8419,8420..8488))
                     /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     8251..8270
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8489..8593)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8619..9074)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8756..8777)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8968..8987
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9201..9223
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9215..9232
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9216..9232
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9242..9261
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9242..9260
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ttaatttaat ttaatgtcct ttaaattttc aaaacatatt ttaatataat tatcatattc
       61 aaagaattgc cataatattc gataaaatga cattattgtt gttttcactt tttttttttt
      121 tttttttttt tttttttttt tcatttataa ttaaaaaggt gtattttaat gtatataaaa
      181 aaggaagtat attatatata taagaataag ataggaataa tacatgagat ggtttattta
      241 tttgtttgtt taaccaattt gagggaaaat atacgtatgt acatactttg ctagtgagct
      301 taaaaaaaac ttttaatatt gtttgtgagg ttaaagaaag catttaatat tgtttgtgag
      361 gttaaagaaa acttttaata ttgtttgtga aaatttttta ctaagaacat aatttttaag
      421 ggatattata aatttattga atttgttatt ataaagtaaa ttttattttc tgcctatttt
      481 atttatgtac aaaaaattgc ggaattgtac ataatataat atgaaaaata tatgacttta
      541 aattgcaatt ggcaaataag gagggggccc cacgccctca aagggtgcca tacactgttg
      601 ggtaatatgg cagcaatcca tatcattaac actatcatta tcttaacatt atcattaaca
      661 ttatcttaac attatcatta acatattgct ttattcgcat taacgttaac atattgcgct
      721 ctttcagccc atgccttgca aataattgca ctttcggaaa tttgaaaaaa tataaagccc
      781 ataaataaag gtggaatata aaacatttat tgcattttcg aagcatataa ataaatatgt
      841 atataattat gcatactata tatatgtata tatttgacta tcttttccaa taaaaaaaaa
      901 aaaaaaaaaa ttatataagg gggcccatat atataggcat attaatagct gctttaacta
      961 aaggggctaa taaatggccc cgtattttgt ttttttttat agctgacaat aaaaataaaa
     1021 agtacgaact ttagtcccat aaattaagaa ctatatatgc ttataaaaag catataaaat
     1081 aaattaatat taatttaaat aaattaatta tatgcaaaaa aatatttaaa aaagaatata
     1141 tataaagaaa aataattata aatattatca ttattattat acatatatat atacacccct
     1201 tttgattata tattttttag agaaaaaaat aaatagtgta ttatttccta taatcgtagt
     1261 taattaattt taataattta caatacacta tatattattt ttattatata taaaatatta
     1321 tttataatta attttttata tttatatgta catattttaa ttgcatcctt tttaataaaa
     1381 atttatttac ccaattgtta tagagcttat ttattatata acttaaaaaa aattaagata
     1441 ttttttattt ttttttaaac atttatttcc aatttatttt taacggtcaa attaaagaaa
     1501 tatatatatt tattgcttaa atttataagt atatatttaa aaaaaaaaat taaagctaaa
     1561 aatatatttc attgtatttt gcatttacgc aattcgctac tattttgcca taagtacatt
     1621 ataatatatg cttgtagcaa attaattaaa taaaaaataa ttacaaaaat ttattatccc
     1681 aataaattat aattgcaatt aaaaaaaaaa cctaggaaaa tgcttaaggc tagtaaagga
     1741 gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg
     1801 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt
     1861 aaatttattt gcactactgg aaaactacct gttccatggc caacacttgt cactactttc
     1921 gcgtatggtc ttcaatgctt tgcgagatac ccagatcata tgaaacagca tgactttttc
     1981 aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg
     2041 aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag
     2101 ttaaaaggta ttgattttaa agaagatgga aacattcttg gacacaaatt ggaatacaac
     2161 tataactcac acaatgtata catcatggca gacaaacaaa agaatggaat caaagttaac
     2221 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa
     2281 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa
     2341 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct tgagtttgta
     2401 acagctgctg ggattacaca tggcatggat gaactataca aaggttcggg agagggcaga
     2461 ggatccctgc taacatgcgg tgatgtcgag gagaatcctg gcccaatggt tggttcgcta
     2521 aactgcatcg tcgctgtgtc ccagaacatg ggcatcggca agaacgggga cctgccctgg
     2581 ccaccgctca ggaacgaatt tagatatttc cagagaatga ccacaacctc ttcagtagaa
     2641 ggtaaacaga atctggtgat tatgggtaag aagacctggt tctccattcc tgagaagaat
     2701 cgacctttaa agggtagaat taatttagtt ctcagcagag aactcaagga acctccacaa
     2761 ggagctcatt ttctttccag aagtctagat gatgccttaa aacttactga acaaccagaa
     2821 ttagcaaata aagtagacat ggtctggata gttggtggca gttctgttta taaggaagcc
     2881 atgaatcacc caggccatct taaactattt gtgacaagga tcatgcaaga ctttgaaagt
     2941 gacacgtttt ttccagaaat tgatttggag aaatataaac ttctgccaga atacccaggt
     3001 gttctctctg atgtccagga ggagaaaggc attaagtaca aatttgaagt atatgagaag
     3061 aatgattaaa ctagtattat taatatatat gaatatatat acatcgttgt atgcgtaata
     3121 tacgtgtaac tgtttttttt aatttttata ttaacttttc cttatacatt ttaatgccat
     3181 taactaataa tattttactt gaaaaaattc taaattttta taactctaat ataaaatctg
     3241 tgaataataa aaaattagca taaagtacga attaaacatc gaatatttgt tttaaagggg
     3301 cacacattat gtgcatataa ttttattctt tgaatttaat tttttgctta taattagatt
     3361 aaattaatat aattttttta ttgaaaataa aaacttattt taaaaaatta acaaatggaa
     3421 gttaatgaaa aatacgaaaa aagtaaaggg ttaattctta tatggtcata taaaaaaaca
     3481 aatgaaataa tgtaaacagt tattcatttt ttaagggtat aaaataagct tataaataat
     3541 gttagcatat taaaaacagt gatataagta tgtgtaaata tgaaagaata taataatgga
     3601 tagttaaaaa aatatatttt aaagggaata ttgacaaatt atatactgaa gcactaaaga
     3661 aaatatcatc tttgtatatt atagcttcac atttccaagc aaatggtgcc acaaatacat
     3721 tcttctctct ccataacttc gtatagtata ccttatacga agttatgcta gcactagtta
     3781 ataaagggca ctttaatttt tatgcataat tagcttatat ttagtgcata tttattttta
     3841 tacatatttt tatatatttt ttgatttaca tggcgtttta tgtttatatg tataacatta
     3901 tatgcatagt atttttttga aaatttaatg gtatttcagt ggatgaaaat attactggtg
     3961 ctttgagggg tgagcattta aagcacaata tctaggatac tacaaaaaat acttaaagta
     4021 tgaatattaa attgtaaact taagcataaa gagctcgaaa agaattaagc ttaactcatc
     4081 tacaagcatc aaattaaatg ttttatgcag tacctttttt taaataatcc attttatttt
     4141 tttatactcc gaggatttta cacctgcact gacactacaa atatagtaat attttatccc
     4201 tacattatat gcattttata atatttttat ttattttaat aatttataca cataagtaca
     4261 tagtatagag ttgaaaaaaa aatacgtgta tttctaaaat aaaatgaaat aaatattatg
     4321 ttattatatt gaatacaata aacattttct ccaaattaaa atttaatatt atataacaaa
     4381 aaaaaataca aaacaccaat taaaaataat aataatttta tccttatttt ttaataactg
     4441 ccatttttaa agtatataat aaaaaataaa ttaaatttta aaaatatttc aaaatataaa
     4501 ataaatatat aagaatatat atttgcttat aaataaataa aaattttata gtcgacaaaa
     4561 tggcaagcaa gtgggatcag aagggtatgg acattgccta tgaggaggcg gccttaggtt
     4621 acaaggaggg tggtgttcct attggcggat gtcttatcaa taacaaagac ggaagtgttc
     4681 tcggtcgtgg tcacaacatg agatttcaaa agggatccgc cacactacat ggtgagatct
     4741 ccactttgga aaactgtggg agattagagg gcaaagtgta caaagatacc actttgtata
     4801 cgacgctgtc tccatgcgac atgtgtacag gtgccatcat catgtatggt attccacgct
     4861 gtgttgtcgg tgagaacgtt aatttcaaaa gtaagggcga gaaatattta caaactagag
     4921 gtcacgaggt tgttgttgtt gacgatgaga ggtgtaaaaa gatcatgaaa caatttatcg
     4981 atgaaagacc tcaggattgg tttgaagata ttggtgaggc ttcggaacca tttaagaacg
     5041 tctacttgct acctcaaaca aaccaattgc tgggtttgta caccatcatc agaaataaga
     5101 atacaactag acctgatttc attttctact ccgatagaat catcagattg ttggttgaag
     5161 aaggtttgaa ccatctacct gtgcaaaagc aaattgtgga aactgacacc aacgaaaact
     5221 tcgaaggtgt ctcattcatg ggtaaaatct gtggtgtttc cattgtcaga gctggtgaat
     5281 cgatggagca aggattaaga gactgttgta ggtctgtgcg tatcggtaaa attttaattc
     5341 aaagggacga ggagactgct ttaccaaagt tattctacga aaaattacca gaggatatat
     5401 ctgaaaggta tgtcttccta ttagacccaa tgctggccac cggtggtagt gctatcatgg
     5461 ctacagaagt cttgattaag agaggtgtta agccagagag aatttacttc ttaaacctaa
     5521 tctgtagtaa ggaagggatt gaaaaatacc atgccgcctt cccagaggtc agaattgtta
     5581 ctggtgccct cgacagaggt ctagatgaaa acaagtatct agttccaggg ttgggtgact
     5641 ttggtgacag atactactgt gtttaagaat tgattattgt tctgtacttc ttttgtgaat
     5701 agtattttta catatgtact gttaatttta aaatgtggaa tgccacgaac atattttttc
     5761 gaacaaaata tatatttttt gtgtcgaaat acataaattt gtgtacctcc tttttgttgt
     5821 atatatgaaa aaggaataaa gtgaatggag acgaattact ctagacccta caacttgtat
     5881 atcactactt tataatacta taatattatt attaacatta aactatagaa gaaatatcaa
     5941 caatacttat gtatttgttc aaactttatc agtatagttt ggtaatcgca tatcatatat
     6001 ttgttatgtg taatatttta tgaatatgcg tctttttttt taactataaa taattcaaaa
     6061 taaaataaca taattaacat ttgcaagcat aattaagatg attactttca catttcatcc
     6121 aaagtggaaa atgaccacca catttgatcg aagcttttac cacttttgat gcattagctc
     6181 tgaaatgtat acaagctttc atttgttaat gttacaaagg gaagttttta tttgggggca
     6241 cattaaaagt gaaaaaatta aaagttaaaa aagttaaaaa atgatcggtg tagttggtca
     6301 tatagacatg ttgttatgca agtctttcaa aacgttcctc ataaaataaa aggagggatc
     6361 tagagagctt tcaattctcc aaacggtatt attggtattt gccgcggtgg agctccagct
     6421 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     6481 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6541 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6601 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6661 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6721 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6781 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6841 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6901 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6961 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7021 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7081 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7141 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7201 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7261 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     7321 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7381 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7441 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7501 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7561 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7621 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7681 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7741 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7801 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7861 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7921 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7981 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     8041 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     8101 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     8161 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     8221 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8281 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8341 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8401 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8461 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8521 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8581 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     8641 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     8701 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     8761 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     8821 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     8881 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     8941 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     9001 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     9061 gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     9121 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     9181 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc
     9241 gtaatacgac tcactatagg gcgaattggg taccactctc gaggaattct ataaaaatag
     9301 cataatataa aatatattat ttaaattgct atacccaata atatacttat tctataatag
     9361 ctatattatt aatttttttt tttttttttt tcatcctctt tatatataca tgtgtatatt
     9421 ttcctcagct ttcaaataac aattcgttat taaagagtca atattatttt aaaaaaccaa
     9481 aaaaaagagt gatgaaatat aataataaac aaattaacta actgagctta aattgtaagg
     9541 aagatgatgt atgtgtccgt cttattacat tttagtcctt tt
//
