LOCUS       sequence-111433-        8913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for Bxb1-mediated recombination of the GFP-Tet1 cDNA into a 
            MIN-tagged locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8913)
  AUTHORS   Mulholland CB, Smets M, Schmidtmann E, Leidescher S, Markaki Y, 
            Hofweber M, Qin W, Manzo M, Kremmer E, Thanisch K, Bauer C, Rombaut 
            P, Herzog F, Leonhardt H, Bultmann S
  TITLE     A modular open platform for systematic functional studies under 
            physiological conditions.
  JOURNAL   Nucleic Acids Res. 2015 May 24. pii: gkv550.
  PUBMED    26007658
REFERENCE   2  (bases 1 to 8913)
  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: "Nucleic 
            Acids Res. 2015 May 24. pii: gkv550."
COMMENT     Sequence Label: attB-GFP-Tet1-Poly(A)
FEATURES             Location/Qualifiers
     source          1..8913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    9..46
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     CDS             join(49..51,52..762)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(49..51,52..762)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(91..112)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(352..371)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     699..720
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      808..817
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    7028..7162
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7052..7071)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7106..7125
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        7298..7345
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7364..8167
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7427..7446)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8037..8056
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(8296..8884)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8376..8395)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gatatcccgg cttgtcgacg acggcggtct ccgtcgtcag gatcatccgt gagcaagggc
       61 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      121 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      181 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
      241 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
      301 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
      361 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
      421 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
      481 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
      541 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
      601 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
      661 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
      721 accgccgccg ggatcactct cggcatggac gagctgtaca agagcggcct gaggagcaga
      781 gcccaggcga gcaacagcgc cgtggacgcc accatgggcg atcgcatgtc tcggtcccgc
      841 cccgcaaagc cttccaaatc agtcaaaaca aagctacaga aaaagaaaga catccagatg
      901 aagacgaaga catcaaagca ggctgtcaga catggggcat cagcgaaggc agtgaacccc
      961 ggaaaaccga agcaattaat taaaagaaga gatggtaaga aggaaacgga agacaaaacc
     1021 ccgacaccgg cgccgagttt tctgacgcga gccggggccg cccgaatgaa tcgggatcga
     1081 aaccaagttc tttttcagaa cccagactcc ttaacttgca acgggtttac aatggctctt
     1141 cgaagaacct ctcttagctg gcgactctcc cagcgcccag tagtcacacc caaacctaag
     1201 aaagtaccac cttcaaagaa gcagtgtaca cataatatcc aggacgagcc gggggtaaag
     1261 cactctgaaa atgattcagt ccccagccaa cacgctaccg tgtccccagg cacagagaat
     1321 ggtgaacaga acaggtgttt agttgaaggg gagagccaag agataaccca gtcttgccct
     1381 gtatttgagg aaagaataga agatacccaa agctgtattt ccgcatctgg gaacctggaa
     1441 gcagaaattt cttggccact ggaagggaca cattgtgaag aactactttc tcatcagaca
     1501 tctgataatg agtgcacttc ccctcaagag tgtgctccat tgccccaaag atcaacttcc
     1561 gaagttacct ctcagaaaaa cactagcaat cagttagcag acttgagctc acaagtagag
     1621 tctatcaagc tgtctgatcc ttctccgaat cctacgggaa gcgaccataa tggctttcca
     1681 gactccagtt ttaggatcgt tcctgaattg gacctgaaaa cctgcatgcc tcttgatgag
     1741 tctgtgtatc ccacagctct gataagattc atcttggcag gctcacagcc ggatgtcttt
     1801 gatactaagc cccaagaaaa aacccttata acaaccccag aacaagtggg aagccaccca
     1861 aatcaagttc tggatgccac ctctgtccta ggacaggcct ttagtactct tccacttcag
     1921 tggggatttt ctggtgctaa cttagttcaa gttgaggccc tgggcaaggg ttcagactca
     1981 ccagaggatc ttggtgctat tactatgcta aaccaacaag aaactgtggc tatggatatg
     2041 gataggaatg ctaccccgga cctgcctatc ttccttccta agcctccaaa cacagttgct
     2101 acctacagta gccctctttt agggcctgag ccccacagct ctacctcctg tgggcttgaa
     2161 gtccaaggtg ctaccccgat tttaactttg gactcaggac acactcccca actcccacca
     2221 aaccctgagt caagttcagt acctctggta atagctgcaa acggtacgcg agctgagaag
     2281 cagtttggta ccagcctttt cccagccgtc ccacaagggt ttacagtcgc agccgagaat
     2341 gaagtccagc atgccccatt agatcttacc cagggttctc aggctgctcc cagcaaactg
     2401 gaaggagaga tttctcgggt cagcatcact ggctcagctg atgtcaaagc cacagcgatg
     2461 tccatgccgg ttacacaggc cagtacctct tctcccccat gtaattctac gccgccaatg
     2521 gtggagagga ggaagcgcaa ggcctgtggg gtctgtgagc cgtgccagca gaaggccaac
     2581 tgtggggagt gcacctactg caagaatcga aagaacagcc accagatctg taagaagagg
     2641 aaatgcgagg tgctcaaaaa gaagccagag gccacctcac aggcacaggt tacaaaagaa
     2701 aacaagaggc cccagaggga aaagaagccc aaagttttaa agacagactt taacaacaaa
     2761 ccagtaaatg gccccaaatc agagtccatg gactgcagta gacgtggtca tggggaagaa
     2821 gagcaaaggt tggatttgat cacacatccc cttgaaaatg taaggaagaa tgcaggcggc
     2881 atgacgggaa ttgaggtgga gaagtgggcg ccaaacaaga aatcacactt agcagagggt
     2941 caagtcaaag gaagctgtga tgccaattta actggggtgg aaaaccccca gccctctgaa
     3001 gatgacaagc agcaaaccaa cccttctccg acatttgccc agaccataag gaacggcatg
     3061 aaaaatgtac actgtttacc cactgacaca caccttccgc tcaacaaatt gaatcatgag
     3121 gaatttagca aggcgttggg aaataattcc tctaaattgc tgactgatcc ctcaaactgc
     3181 aaggatgcca tgagtgtcac cacctccggg ggggagtgtg atcatctcaa ggggccaagg
     3241 aacaccctgc tcttccagaa gcctggcttg aactgcaggt ctggtgcaga acccaccatc
     3301 tttaataatc accctaatac acacagcgct gggagtcgac ctcaccctcc tgagaaggta
     3361 cccaacaaag aacccaaaga tggctctccg gttcagccaa gtcttctatc cctaatgaag
     3421 gacaggagat taacgctgga acaagtggta gccatagaag ccctgactca gctctcagaa
     3481 gcgccctcag agagctcctc cccatccaag ccggagaagg atgaagaagc gcatcagaaa
     3541 acggctagtc tgcttaacag ttgcaaggct atccttcatt cggtgagaaa agacctccaa
     3601 gacccaaacg ttcaaggaaa gggccttcac catgacactg ttgtctttaa tggccagaac
     3661 agaacattca agtcacctga tagtttcgct actaaccaag cacttataaa atcacaagga
     3721 tatccaagtt cacctacagc tgagaagaaa ggcgctgctg gtgggagagc tccctttgat
     3781 ggttttgaga attcacatcc attgcccata gaaagtcaca atcttgaaaa ctgcagtcag
     3841 gtactaagct gcgatcagaa tttgagttcc catgacccat catgccagga tgcaccctac
     3901 agtcaaattg aagaagatgt cgcagcacaa ttgacccaac ttgcatccac gattaatcac
     3961 atcaacgccg aggtcaggaa tgctgaaagt acaccagaaa gccttgttgc aaagaacaca
     4021 aagcaaaaac acagtcagga gaaacgcatg gtacatcaga agccaccctc tagtacacaa
     4081 actaaaccca gtgtaccatc agcaaagcca aagaaggccc agaaaaaggc aagagcgacc
     4141 ccacacgcaa acaagcggaa aaagaaaccc ccagcccgaa gctctcaaga aaatgatcag
     4201 aagaagcaag agcagctagc tatagagtat agtaaaatgc atgacatttg gatgtcatct
     4261 aagtttcaaa ggtttggcca gtctagtcca cgtagttttc ctgttctgtt gagaaatatt
     4321 cctgtcttta accaaatatt aaagcccgtg actcagagta aaacaccctc acaacataat
     4381 gagttatttc ctcctatcaa tcagataaaa ttcacaagaa accctgaatt ggcaaaggag
     4441 aaagtgaagg ttgaaccatc agattctctg ccaacatgcc aattcaagac agaatccggt
     4501 gggcagacat ttgcagagcc ggcagataat tctcagggac agcccatggt gagtgtcaat
     4561 caggaagctc accctctgcc ccagtcccca ccatctaacc agtgtgctaa tataatggct
     4621 ggtgctgccc aaacacagtt tcatctaggc gctcaagaga acctggtgca tcagatacca
     4681 ccaccgacgt tgcctggtac ctcccctgac acactcttgc cagacccagc atcaatcctc
     4741 aggaaaggga aagtgttaca tttcgatgga attacagttg ttacggagaa gcgtgaagct
     4801 caaacatcaa gcaatggacc actgggtccc accacagaca gtgcacagtc agaatttaaa
     4861 gagagcatca tggacttact tagcaagcct gcaaaaaacc tgatagctgg attgaaggaa
     4921 caggaagctg caccctgtga ctgtgatgga ggtacacaaa aagaaaaagg cccatattat
     4981 acacaccttg gggcaggacc aagtgtggct gctgtcaggg agctcatgga gactaggttt
     5041 ggccagaagg ggaaggcaat ccggattgag aagatagtgt tcacggggaa ggaagggaag
     5101 agctctcagg gctgcccggt cgccaagtgg gtgatcagaa gaagtggtcc tgaagagaag
     5161 cttatttgtt tggttcgtga gcgtgtagac catcactgtt cgacggctgt gatagttgtc
     5221 cttatcctgc tgtgggaagg tatccctcgc ctgatggctg accgcctgta caaagagctc
     5281 actgagaact tgaggtccta cagcggacat cccacagacc gaagatgtac cctcaacaaa
     5341 aagcgtacct gcacctgtca aggcatcgac ccaaaaacct gcggagcgtc cttctccttt
     5401 ggctgttcgt ggagcatgta tttcaacggc tgtaagtttg ggaggagtga aaaccccaga
     5461 aaattcagac ttgctccaaa ctacccctta cataactact ataagagaat tactggaatg
     5521 agttctgaag gaagtgacgt gaaaaccggg tggatcattc cagaccgcaa gaccctcata
     5581 agcagagagg aaaaacagct tgaaaagaat ttacaagaat tggctacagt attagctcca
     5641 ctttacaagc agatggctcc agttgcttat caaaatcagg tggaatatga agaagttgct
     5701 ggagactgtc gacttggaaa tgaagagggg cgtcctttct ctggtgtcac ctgttgcatg
     5761 gatttttgtg cccattctca caaggacatt cacaacatgc acaacggaag caccgtggtg
     5821 tgtacgttga ttcgagcaga tggccgtgac acaaattgtc ccgaggatga acaactccac
     5881 gtcctgccac tataccggct tgcagacact gatgaatttg gctccgtgga agggatgaag
     5941 gccaaaatca aatctggggc catccaagtc aatgggccaa ccaggaagag gcgactacgt
     6001 tttactgagc ctgttcctcg atgtgggaag agggccaaaa tgaagcagaa ccacaataaa
     6061 tcaggtactg ctggtctcag aaggaagagg atctctgcca gcccaaaggg agccccaggt
     6121 tcacacaaca ctaagagctt ttcatcagcc tcatctactt ctcacctagt gaaagacgaa
     6181 tctacagact tctgtcccct gcaggcttcc tccgcagaaa catctacctg tacgtacagt
     6241 aaaacagcct caggtgggtt tgcagaaaca agtagtattc tccactgcac aatgccttct
     6301 ggagcacaca gtggtgctaa tgcagctgct ggggaatgta ctggaacggt gcagcctgcc
     6361 gaggtggctg ctcatcctca ccagtctctt cccacagccg attctcccgt tcatgctgag
     6421 cctctcacta gtccatctga gcagctaact tctaaccagt caaaccagca gctccctctc
     6481 ctcagcaatt ctcagaaact ggcttcctgt caggtggaag atgagcggca ccctgaagcg
     6541 gatgagcctc agcaccccga ggacgataac ttgcctcaac ttgatgaatt ctggtcagac
     6601 agtgaggaga tctacgccga tccttccttt ggtggcgtgg cgatagcacc cattcacggc
     6661 tcggtgctca ttgagtgcgc tcggaaggag cttcatgcta ccacctcttt gcgctccccc
     6721 aaacgagggg tcccttttcg tgtgtccctt gtattctacc agcacaaaag cctaaacaag
     6781 cctaatcatg gttttgatat caacaaaatt aagtgtaaat gcaaaaaagt aacgaaaaaa
     6841 aagcccgcag accgggagtg tcctgatgta tcccccgaag ccaatttatc acaccaaatt
     6901 ccttctcgag ttgcatcaac cttaacccga gacaatgttg ttaccgtgtc cccatactct
     6961 ctcactcatg ttgcgggacc ctacaatcgt tgggtctaag cggccgcagt taacgaattc
     7021 tctagaggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     7081 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     7141 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     7201 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     7261 ttatgatcct gcaagcctcg tcgccgcggt ttattctgtt gacaattaat catcggcata
     7321 gtatatcggc atagtataat acgacaaggt gaggaactaa accatgggat cggccattga
     7381 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     7441 ctgggcacaa cagacgatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     7501 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     7561 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     7621 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     7681 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     7741 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     7801 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     7861 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     7921 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     7981 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     8041 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     8101 ttacggtatc gccgcccccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     8161 cttctgagcg ggactctggg gttcgaataa agaccgacca agcgacgtct gagagctccc
     8221 tggcgaattc ggtaccaata aaagagcttt attttcatga tctgtgtgtt ggtttttggc
     8281 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     8341 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     8401 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     8461 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     8521 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     8581 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     8641 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     8701 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     8761 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     8821 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     8881 tcaagaagat cctttgatct tttctagtgt gcg
//
