LOCUS       sequence-111431-        8253 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for Bxb1-mediated recombination of the GFP-Tet1d834-1053 
            cDNA into a MIN-tagged locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8253)
  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 8253)
  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-Tet1d834-1053-Poly(A)
FEATURES             Location/Qualifiers
     source          1..8253
                     /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    6368..6502
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6392..6411)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6446..6465
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        6638..6685
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6704..7507
                     /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(6767..6786)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7377..7396
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(7636..8224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7716..7735)
                     /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 acacagcagt acaccagaaa gccttgttgc aaagaacaca
     3361 aagcaaaaac acagtcagga gaaacgcatg gtacatcaga agccaccctc tagtacacaa
     3421 actaaaccca gtgtaccatc agcaaagcca aagaaggccc agaaaaaggc aagagcgacc
     3481 ccacacgcaa acaagcggaa aaagaaaccc ccagcccgaa gctctcaaga aaatgatcag
     3541 aagaagcaag agcagctagc tatagagtat agtaaaatgc atgacatttg gatgtcatct
     3601 aagtttcaaa ggtttggcca gtctagtcca cgtagttttc ctgttctgtt gagaaatatt
     3661 cctgtcttta accaaatatt aaagcccgtg actcagagta aaacaccctc acaacataat
     3721 gagttatttc ctcctatcaa tcagataaaa ttcacaagaa accctgaatt ggcaaaggag
     3781 aaagtgaagg ttgaaccatc agattctctg ccaacatgcc aattcaagac agaatccggt
     3841 gggcagacat ttgcagagcc ggcagataat tctcagggac agcccatggt gagtgtcaat
     3901 caggaagctc accctctgcc ccagtcccca ccatctaacc agtgtgctaa tataatggct
     3961 ggtgctgccc aaacacagtt tcatctaggc gctcaagaga acctggtgca tcagatacca
     4021 ccaccgacgt tgcctggtac ctcccctgac acactcttgc cagacccagc atcaatcctc
     4081 aggaaaggga aagtgttaca tttcgatgga attacagttg ttacggagaa gcgtgaagct
     4141 caaacatcaa gcaatggacc actgggtccc accacagaca gtgcacagtc agaatttaaa
     4201 gagagcatca tggacttact tagcaagcct gcaaaaaacc tgatagctgg attgaaggaa
     4261 caggaagctg caccctgtga ctgtgatgga ggtacacaaa aagaaaaagg cccatattat
     4321 acacaccttg gggcaggacc aagtgtggct gctgtcaggg agctcatgga gactaggttt
     4381 ggccagaagg ggaaggcaat ccggattgag aagatagtgt tcacggggaa ggaagggaag
     4441 agctctcagg gctgcccggt cgccaagtgg gtgatcagaa gaagtggtcc tgaagagaag
     4501 cttatttgtt tggttcgtga gcgtgtagac catcactgtt cgacggctgt gatagttgtc
     4561 cttatcctgc tgtgggaagg tatccctcgc ctgatggctg accgcctgta caaagagctc
     4621 actgagaact tgaggtccta cagcggacat cccacagacc gaagatgtac cctcaacaaa
     4681 aagcgtacct gcacctgtca aggcatcgac ccaaaaacct gcggagcgtc cttctccttt
     4741 ggctgttcgt ggagcatgta tttcaacggc tgtaagtttg ggaggagtga aaaccccaga
     4801 aaattcagac ttgctccaaa ctacccctta cataactact ataagagaat tactggaatg
     4861 agttctgaag gaagtgacgt gaaaaccggg tggatcattc cagaccgcaa gaccctcata
     4921 agcagagagg aaaaacagct tgaaaagaat ttacaagaat tggctacagt attagctcca
     4981 ctttacaagc agatggctcc agttgcttat caaaatcagg tggaatatga agaagttgct
     5041 ggagactgtc gacttggaaa tgaagagggg cgtcctttct ctggtgtcac ctgttgcatg
     5101 gatttttgtg cccattctca caaggacatt cacaacatgc acaacggaag caccgtggtg
     5161 tgtacgttga ttcgagcaga tggccgtgac acaaattgtc ccgaggatga acaactccac
     5221 gtcctgccac tataccggct tgcagacact gatgaatttg gctccgtgga agggatgaag
     5281 gccaaaatca aatctggggc catccaagtc aatgggccaa ccaggaagag gcgactacgt
     5341 tttactgagc ctgttcctcg atgtgggaag agggccaaaa tgaagcagaa ccacaataaa
     5401 tcaggtactg ctggtctcag aaggaagagg atctctgcca gcccaaaggg agccccaggt
     5461 tcacacaaca ctaagagctt ttcatcagcc tcatctactt ctcacctagt gaaagacgaa
     5521 tctacagact tctgtcccct gcaggcttcc tccgcagaaa catctacctg tacgtacagt
     5581 aaaacagcct caggtgggtt tgcagaaaca agtagtattc tccactgcac aatgccttct
     5641 ggagcacaca gtggtgctaa tgcagctgct ggggaatgta ctggaacggt gcagcctgcc
     5701 gaggtggctg ctcatcctca ccagtctctt cccacagccg attctcccgt tcatgctgag
     5761 cctctcacta gtccatctga gcagctaact tctaaccagt caaaccagca gctccctctc
     5821 ctcagcaatt ctcagaaact ggcttcctgt caggtggaag atgagcggca ccctgaagcg
     5881 gatgagcctc agcaccccga ggacgataac ttgcctcaac ttgatgaatt ctggtcagac
     5941 agtgaggaga tctacgccga tccttccttt ggtggcgtgg cgatagcacc cattcacggc
     6001 tcggtgctca ttgagtgcgc tcggaaggag cttcatgcta ccacctcttt gcgctccccc
     6061 aaacgagggg tcccttttcg tgtgtccctt gtattctacc agcacaaaag cctaaacaag
     6121 cctaatcatg gttttgatat caacaaaatt aagtgtaaat gcaaaaaagt aacgaaaaaa
     6181 aagcccgcag accgggagtg tcctgatgta tcccccgaag ccaatttatc acaccaaatt
     6241 ccttctcgag ttgcatcaac cttaacccga gacaatgttg ttaccgtgtc cccatactct
     6301 ctcactcatg ttgcgggacc ctacaatcgt tgggtctaag cggccgcagt taacgaattc
     6361 tctagaggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     6421 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     6481 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     6541 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     6601 ttatgatcct gcaagcctcg tcgccgcggt ttattctgtt gacaattaat catcggcata
     6661 gtatatcggc atagtataat acgacaaggt gaggaactaa accatgggat cggccattga
     6721 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     6781 ctgggcacaa cagacgatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     6841 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     6901 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     6961 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     7021 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     7081 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     7141 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     7201 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     7261 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     7321 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     7381 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     7441 ttacggtatc gccgcccccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     7501 cttctgagcg ggactctggg gttcgaataa agaccgacca agcgacgtct gagagctccc
     7561 tggcgaattc ggtaccaata aaagagcttt attttcatga tctgtgtgtt ggtttttggc
     7621 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     7681 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     7741 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     7801 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     7861 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     7921 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7981 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     8041 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     8101 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     8161 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     8221 tcaagaagat cctttgatct tttctagtgt gcg
//
