LOCUS       sequence-111432-        7323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for Bxb1-mediated recombination of the GFP-Tet1d1054-1363 
            cDNA into a MIN-tagged locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7323)
  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 7323)
  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-Tet1d834-1363-Poly(A)
FEATURES             Location/Qualifiers
     source          1..7323
                     /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    5438..5572
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5462..5481)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5516..5535
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5708..5755
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5774..6577
                     /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(5837..5856)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6447..6466
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(6706..7294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6786..6805)
                     /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 acacagcgaa caggaagctg caccctgtga ctgtgatgga
     3361 ggtacacaaa aagaaaaagg cccatattat acacaccttg gggcaggacc aagtgtggct
     3421 gctgtcaggg agctcatgga gactaggttt ggccagaagg ggaaggcaat ccggattgag
     3481 aagatagtgt tcacggggaa ggaagggaag agctctcagg gctgcccggt cgccaagtgg
     3541 gtgatcagaa gaagtggtcc tgaagagaag cttatttgtt tggttcgtga gcgtgtagac
     3601 catcactgtt cgacggctgt gatagttgtc cttatcctgc tgtgggaagg tatccctcgc
     3661 ctgatggctg accgcctgta caaagagctc actgagaact tgaggtccta cagcggacat
     3721 cccacagacc gaagatgtac cctcaacaaa aagcgtacct gcacctgtca aggcatcgac
     3781 ccaaaaacct gcggagcgtc cttctccttt ggctgttcgt ggagcatgta tttcaacggc
     3841 tgtaagtttg ggaggagtga aaaccccaga aaattcagac ttgctccaaa ctacccctta
     3901 cataactact ataagagaat tactggaatg agttctgaag gaagtgacgt gaaaaccggg
     3961 tggatcattc cagaccgcaa gaccctcata agcagagagg aaaaacagct tgaaaagaat
     4021 ttacaagaat tggctacagt attagctcca ctttacaagc agatggctcc agttgcttat
     4081 caaaatcagg tggaatatga agaagttgct ggagactgtc gacttggaaa tgaagagggg
     4141 cgtcctttct ctggtgtcac ctgttgcatg gatttttgtg cccattctca caaggacatt
     4201 cacaacatgc acaacggaag caccgtggtg tgtacgttga ttcgagcaga tggccgtgac
     4261 acaaattgtc ccgaggatga acaactccac gtcctgccac tataccggct tgcagacact
     4321 gatgaatttg gctccgtgga agggatgaag gccaaaatca aatctggggc catccaagtc
     4381 aatgggccaa ccaggaagag gcgactacgt tttactgagc ctgttcctcg atgtgggaag
     4441 agggccaaaa tgaagcagaa ccacaataaa tcaggtactg ctggtctcag aaggaagagg
     4501 atctctgcca gcccaaaggg agccccaggt tcacacaaca ctaagagctt ttcatcagcc
     4561 tcatctactt ctcacctagt gaaagacgaa tctacagact tctgtcccct gcaggcttcc
     4621 tccgcagaaa catctacctg tacgtacagt aaaacagcct caggtgggtt tgcagaaaca
     4681 agtagtattc tccactgcac aatgccttct ggagcacaca gtggtgctaa tgcagctgct
     4741 ggggaatgta ctggaacggt gcagcctgcc gaggtggctg ctcatcctca ccagtctctt
     4801 cccacagccg attctcccgt tcatgctgag cctctcacta gtccatctga gcagctaact
     4861 tctaaccagt caaaccagca gctccctctc ctcagcaatt ctcagaaact ggcttcctgt
     4921 caggtggaag atgagcggca ccctgaagcg gatgagcctc agcaccccga ggacgataac
     4981 ttgcctcaac ttgatgaatt ctggtcagac agtgaggaga tctacgccga tccttccttt
     5041 ggtggcgtgg cgatagcacc cattcacggc tcggtgctca ttgagtgcgc tcggaaggag
     5101 cttcatgcta ccacctcttt gcgctccccc aaacgagggg tcccttttcg tgtgtccctt
     5161 gtattctacc agcacaaaag cctaaacaag cctaatcatg gttttgatat caacaaaatt
     5221 aagtgtaaat gcaaaaaagt aacgaaaaaa aagcccgcag accgggagtg tcctgatgta
     5281 tcccccgaag ccaatttatc acaccaaatt ccttctcgag ttgcatcaac cttaacccga
     5341 gacaatgttg ttaccgtgtc cccatactct ctcactcatg ttgcgggacc ctacaatcgt
     5401 tgggtctaag cggccgcagt taacgaattc tctagaggat ccagacatga taagatacat
     5461 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     5521 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     5581 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     5641 gtaaaacctc tacaaatgtg gtatggctga ttatgatcct gcaagcctcg tcgccgcggt
     5701 ttattctgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     5761 gaggaactaa accatgggat cggccattga acaagatgga ttgcacgcag gttctccggc
     5821 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacgatcg gctgctctga
     5881 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     5941 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     6001 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     6061 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     6121 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     6181 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     6241 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     6301 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     6361 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     6421 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     6481 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgcccccg attcgcagcg
     6541 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaataa
     6601 agaccgacca agcgacgtct gagagctccc tggcgaattc ggtaccaata aaagagcttt
     6661 attttcatga tctgtgtgtt ggtttttggc cgcgttgctg gcgtttttcc ataggctccg
     6721 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6781 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6841 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6901 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6961 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7021 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7081 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7141 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7201 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7261 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctagtgt
     7321 gcg
//
