LOCUS       sequence-477249-        5039 bp    DNA     linear   UNA 08-JUL-2025
DEFINITION  N_terminal segment of the blue_light_controlled split PS_intein tTA 
            gene expression system, co_expressing mTagBFP2 under the BIRC5 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5039)
  TITLE     Photoswitchable intein for light control of covalent protein binding
            and cleavage
REFERENCE   2  (bases 1 to 5039)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pBIRC5-Split PS Intein-N tTA
FEATURES             Location/Qualifiers
     source          1..5039
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             938..991
                     /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"
     CDS             995..1693
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     polyA_signal    1816..1937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1853..1872)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1907..1926
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1944..2399)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2081..2102)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2293..2312
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2426..2530
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2532..2889
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2532..2723
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2555..2575)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2740..2875
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2924..3718
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2924..3718
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2978..2997)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3588..3607
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3906..3925)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3950..3997
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4326..4914
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4815..4834
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gatgcttttt gcagaggtgg caccctgtaa agctctcctg tctgactttt tttttttttt
       61 agactgagtt ttgctcttgt tgcctaggct ggagtgcaat ggcacaatct cagctcactg
      121 caccctctgc ctcccgggtt caagcgattc tcctgcctca gcctcccgag tagttgggat
      181 tacaggcatg caccaccacg cccagctaat ttttgtattt ttagtagaga caaggtttca
      241 ccgtgatggc caggctggtc ttgaactcca ggactcaagt gatgctcctg cctaggcctc
      301 tcaaagtgtt gggattacag gcgtgagcca ctgcacccgg cctgcacgcg ttctttgaaa
      361 gcagtcgagg gggcgctagg tgtgggcagg gacgagctgg cgcggcgtcg ctgggtgcac
      421 cgcgaccacg ggcagagcca cgcggcggga ggactacaac tcccggcaca ccccgcgccg
      481 ccccgcctct actcccagaa ggccgcgggg ggtggaccgc ctaagagggc gtgcgctccc
      541 gacatgcccc gcggcgcgcc attaaccgcc agatttgaat cgccggaccc gttggcagag
      601 gtggcggcgg cgagagctgg tttagtgaac cgtcagatcc gctagcacca tgagcggcta
      661 ttgtctggat ctgaaaaccc aggttcagac accgcagggt atgaaagaaa tttcaaatat
      721 tcaggtgggt gatctggttc tgagcaatac tggttataat gaagtgctga atgtgtttcc
      781 gaaaagcaaa aaaaaaagct ataaaatcac cctggaagat ggcaaagaaa tcatttgtag
      841 cgaagaacac ctgtttccga cccagactgg tgaaatgaat attagcggtg gtctgaaaga
      901 aggtatgtgc ctgtatgtta aagaattcgg tagcggcgag ggcagaggaa gtctgctaac
      961 atgcggtgat gtcgaggaga atcctggccc aggaagcgag ctgattaagg agaacatgca
     1021 catgaagctg tacatggagg gcaccgtgga caaccatcac ttcaagtgca catccgaggg
     1081 cgaaggcaag ccctacgagg gcacccagac catgagaatc aaggtggtcg agggcggccc
     1141 tctccccttc gccttcgaca tcctggctac tagcttcctc tacggcagca agaccttcat
     1201 caaccacacc cagggcatcc ccgacttctt caagcagtcc ttccctgagg gcttcacatg
     1261 ggagagagtc accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct
     1321 ccaggacggc tgcctcatct acaacgtcaa gatcagaggg gtgaacttca catccaacgg
     1381 ccctgtgatg cagaagaaaa cactcggctg ggaggccttc accgagacgc tgtaccccgc
     1441 tgacggcggc ctggaaggca gaaacgacat ggccctgaag ctcgtgggcg ggagccatct
     1501 gatcgcaaac gccaagacca catatagatc caagaaaccc gctaagaacc tcaagatgcc
     1561 tggcgtctac tatgtggact acagactgga aagaatcaag gaggccaaca acgagaccta
     1621 cgtcgagcag cacgaggtgg cagtggccag atactgcgac ctccctagca aactggggca
     1681 caagcttaat taaagcggcc gcgactctag atcataatca gccataccac atttgtagag
     1741 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     1801 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1861 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1921 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     1981 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     2041 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     2101 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     2161 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     2221 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     2281 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     2341 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     2401 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2461 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2521 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     2581 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2641 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2701 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     2761 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     2821 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     2881 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     2941 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     3001 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     3061 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     3121 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     3181 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     3241 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     3301 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     3361 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     3421 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     3481 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     3541 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     3601 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     3661 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     3721 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     3781 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     3841 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     3901 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     3961 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     4021 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     4081 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     4141 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     4201 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4261 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4321 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4381 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4441 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4501 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4561 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4621 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4681 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4741 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4801 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4861 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4921 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     4981 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaat
//
