LOCUS       sequence-481812-        6088 bp    DNA     circular SYN 08-AUG-2025
DEFINITION  N_terminal segment of the blue_light_controlled split PS_intein tTA 
            gene expression system, co_expressing mTagBFP2 under the hTERT 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6088)
  TITLE     Photoswitchable intein for light control of covalent protein binding
            and cleavage
REFERENCE   2  (bases 1 to 6088)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: phTERT-Split PS Intein-N tTA
FEATURES             Location/Qualifiers
     source          1..6088
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1987..2040
                     /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             2044..2742
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     polyA_signal    2865..2986
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2902..2921)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2956..2975
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2993..3448)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3130..3151)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3342..3361
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3475..3579
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3581..3938
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3581..3772
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3604..3624)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3789..3924
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3973..4767
                     /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             3973..4767
                     /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(4027..4046)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4637..4656
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4955..4974)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4999..5046
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5375..5963
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5864..5883
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcatcagctt ttcaaagaca cactaactgc acccataata ctggggtgtc ttctgggtat
       61 cagcgatctt cattgaatgc cgggaggcgt ttcctcgcca tgcacatggt gttaattact
      121 ccagcataat cttctgcttc catttcttct cttccctctt ttaaaattgt gttttctatg
      181 ttggcttctc tgcagagaac cagtgtaagc tacaacttaa cttttgttgg aacaaatttt
      241 ccaaaccgcc cctttgccct agtggcagag acaattcaca aacacagccc tttaaaaagg
      301 cttagggatc actaagggga tttctagaag agcgacccgt aatcctaagt atttacaaga
      361 cgaggctaac ctccagcgag cgtgacagcc cagggagggt gcgaggcctg ttcaaatgct
      421 agctccataa ataaagcaat ttcctccggc agtttctgaa agtaggaaag gttacattta
      481 aggttgcgtt tgttagcatt tcagtgtttg ccgacctcag ctacagcatc cctgcaaggc
      541 ctcgggagac ccagaagttt ctcgcccctt agatccaaac ttgagcaacc cggagtctgg
      601 attcctggga agtcctcagc tgtcctgcgg ttgtgccggg gccccaggtc tggaggggac
      661 cagtggccgt gtggcttcta ctgctgggct ggaagtcggg cctcctagct ctgcagtccg
      721 aggcttggag ccaggtgcct ggaccccgag gctgccctcc accctgtgcg ggcgggatgt
      781 gaccagatgt tggcctcatc tgccagacag agtgccgggg cccagggtca aggccgttgt
      841 ggctggtgtg aggcgcccgg tgcgcggcca gcaggagcgc ctggctccat ttcccaccct
      901 ttctcgacgg gaccgccccg gtgggtgatt aacagatttg gggtggtttg ctcatggtgg
      961 ggacccctcg ccgcctgaga acctgcaaag agaaatgacg ggcctgtgtc aaggagccca
     1021 agtcgcgggg aagtgttgca gggaggcact ccgggaggtc ccgcgtgccc gtccagggag
     1081 caatgcgtcc tcgggttcgt ccccagccgc gtctacgcgc ctccgtcctc cccttcacgt
     1141 ccggcattcg tggtgcccgg agcccgacgc cccgcgtccg gacctggagg cagccctggg
     1201 tctccggatc aggccagcgg ccaaagggtc gccgcacgca cctgttccca gggcctccac
     1261 atcatggccc ctccctcggg ttaccccaca gcctaggccg attcgacctc tctccgctgg
     1321 ggccctcgct ggcgtccctg caccctggga gcgcgagcgg cgcgcgggcg gggaagcgcg
     1381 gcccagaccc ccgggtccgc ccggagcagc tgcgctgtcg gggccaggcc gggctcccag
     1441 tggattcgcg ggcacagacg cccaggaccg cgcttcccac gtggcggagg gactggggac
     1501 ccgggcaccc gtcctgcccc ttcaccttcc agctccgcct cctccgcgcg gaccccgccc
     1561 cgtcccgacc cctcccgggt ccccggccca gccccctccg ggccctccca gcccctcccc
     1621 ttcctttccg cggccccgcc ctctcctcgc ggcgcgagtt tagagctggt ttagtgaacc
     1681 gtcagatccg ctagcaccat gagcggctat tgtctggatc tgaaaaccca ggttcagaca
     1741 ccgcagggta tgaaagaaat ttcaaatatt caggtgggtg atctggttct gagcaatact
     1801 ggttataatg aagtgctgaa tgtgtttccg aaaagcaaaa aaaaaagcta taaaatcacc
     1861 ctggaagatg gcaaagaaat catttgtagc gaagaacacc tgtttccgac ccagactggt
     1921 gaaatgaata ttagcggtgg tctgaaagaa ggtatgtgcc tgtatgttaa agaattcggt
     1981 agcggcgagg gcagaggaag tctgctaaca tgcggtgatg tcgaggagaa tcctggccca
     2041 ggaagcgagc tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtggac
     2101 aaccatcact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc
     2161 atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctact
     2221 agcttcctct acggcagcaa gaccttcatc aaccacaccc agggcatccc cgacttcttc
     2281 aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc
     2341 gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag
     2401 atcagagggg tgaacttcac atccaacggc cctgtgatgc agaagaaaac actcggctgg
     2461 gaggccttca ccgagacgct gtaccccgct gacggcggcc tggaaggcag aaacgacatg
     2521 gccctgaagc tcgtgggcgg gagccatctg atcgcaaacg ccaagaccac atatagatcc
     2581 aagaaacccg ctaagaacct caagatgcct ggcgtctact atgtggacta cagactggaa
     2641 agaatcaagg aggccaacaa cgagacctac gtcgagcagc acgaggtggc agtggccaga
     2701 tactgcgacc tccctagcaa actggggcac aagcttaatt aaagcggccg cgactctaga
     2761 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     2821 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     2881 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2941 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     3001 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3061 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3121 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3181 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3241 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3301 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3361 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3421 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3481 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3541 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     3601 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3661 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3721 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3781 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3841 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3901 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     3961 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4021 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4081 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4141 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4201 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4261 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4321 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4381 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4441 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4501 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4561 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4621 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4681 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4741 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4801 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4861 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4921 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     4981 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     5041 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     5101 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5161 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5221 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5281 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5341 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5401 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5461 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5521 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5581 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5641 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5701 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5761 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5821 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5881 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5941 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6001 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6061 gtattaccgc catgcattag ttattaat
//
