LOCUS       sequence-210240-        6655 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PLC pleckstrin homology domain linked to FAP for CALI validation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6655)
  AUTHORS   He J, Wang Y, Missinato MA, Onuoha E, Perkins LA, Watkins SC, St 
            Croix CM, Tsang M, Bruchez MP
  TITLE     A genetically targetable near-infrared photosensitizer.
  JOURNAL   Nat Methods. 2016 Mar;13(3):263-268. doi: 10.1038/nmeth.3735. Epub 
            2016 Jan 25.
  PUBMED    26808669
REFERENCE   2  (bases 1 to 6655)
  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"; doi: 
            "10.1038/nmeth.3735"; journalName: "Nat Methods"; date: "2016-03"; 
            volume: "13"; issue: "3"; pages: "263-268"
COMMENT     Sequence Label: PH-dL5**
FEATURES             Location/Qualifiers
     source          1..6655
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1471..2190
                     /codon_start=1
                     /product="fluorogen-activating protein composed of a dimer 
                     of the L5-MG single-chain antibody fragment (Szent-Gyorgyi 
                     et al., 2008)"
                     /label=FAP-alpha-2
                     /note="binds malachite green derivatives to produce red 
                     fluorescence"
                     /translation="QAVVTQEPSVTVSPGGTVILTCGSGTGAVTSGHYANWFQQKPGQA
                     PRALIFDTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSLSDVDGYLFGGGT
                     QLTVLSGGGGSGGGGSGGGGSGGGGSQAVVTQEPSVTVSPGGTVILTCGSGTGAVTSGH
                     YANWFQQKPGQAPRALIFDTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSL
                     SDVDGYLFGGGTQLTVLS"
     primer_bind     complement(2246..2263)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2252..2476
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2522..2950
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2609..2628)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2819..2840
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2959..2979)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2964..3293
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3144..3279
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3206..3225
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3360..4154
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3414..3433)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4024..4043
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4328..4449
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4365..4384)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4419..4438
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4498..4514)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4498..4514)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4511..4533)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4522..4538
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4546..4552,4553..4570,4571..4576))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4591..4612
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4729..4746)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4900..5488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4980..4999)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5659..6450,6451..6519))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6282..6301
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6520..6624)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      901 gcgtttaaac ttaagcttat ggactcgggc cgggacttcc tgaccctgca cggcctacag
      961 gatgatgagg atctacaggc gctgctgaag ggcagccagc tcctgaaggt gaagtccagc
     1021 tcatggagga gagagcgctt ctacaagttg caggaggact gcaagaccat ctggcaggag
     1081 tcccgcaagg tcatgcggac cccggagtcc cagctgttct ccatcgagga cattcaggag
     1141 gtgcgaatgg ggcaccgcac ggagggtctg gagaagttcg cccgtgatgt gcccgaggac
     1201 cgctgcttct ccattgtctt caaggaccag cgcaatacac tagacctcat cgccccatcg
     1261 ccagctgatg cccagcactg ggtgctgggg ctgcacaaga tcatccacca ctcaggctcc
     1321 atggaccagc gtcagaagct acagcactgg attcactcct gcttgcgaaa agctgacaaa
     1381 aacaaggaca acaagatgag cttcaaggag ctgcagaact tcctgaagca tatgggaggc
     1441 ggtgggtctg gtggcggtgg cagcaccggt caggccgtcg ttacccaaga acctagtgtt
     1501 accgttagcc caggtggtac tgttatactt acttgtggaa gtggtacggg tgccgtcaca
     1561 tctggtcatt atgcaaattg gtttcaacaa aaaccaggac aagctccaag agctttgatt
     1621 tttgatactg ataagaagta ttcttggacc ccaggtagat tttctggatc tttgctggga
     1681 gcaaaggcag ctttgacaat atcagatgct cagcctgagg acgaagccga gtattactgt
     1741 tctcttagcg acgtggatgg ctacttgttt ggcggtggaa cacaactgac ggttctgtcc
     1801 ggtggtggcg gctctggtgg cggtggcagc ggcggtggtg gttccggagg cggcggttct
     1861 caggctgtgg tgactcagga gccgtcagtg actgtgtccc caggagggac agtcattctc
     1921 acttgtggct ccggcactgg agctgtcacc agtggtcatt atgccaactg gttccagcag
     1981 aagcctggcc aagcccccag ggcacttata tttgacaccg acaagaagta ttcctggacc
     2041 cctggccgat tctcaggctc cctccttggg gccaaggctg ccctgaccat ctcggatgcg
     2101 cagcctgaag atgaggctga gtattactgt tcgctctccg acgttgacgg ttatctgttc
     2161 ggaggaggca cccagctgac cgtcctctcc taatagctcg aggcagctgt cgactctaga
     2221 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2281 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2341 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2401 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2461 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     2521 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2581 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2641 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2701 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2761 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2821 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2881 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2941 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3001 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3061 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3121 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3181 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3241 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3301 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     3361 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3421 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3481 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3541 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3601 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3661 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3721 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3781 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3841 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     3901 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     3961 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4021 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4081 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4141 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4201 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4261 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4321 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4381 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4441 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4501 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4561 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4621 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4681 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4741 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4801 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4861 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4921 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4981 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5041 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5101 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5161 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5221 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5281 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5341 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5401 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5461 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5521 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5581 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5641 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5701 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5761 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5821 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5881 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5941 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6001 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6061 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6121 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6181 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6241 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6301 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6361 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6421 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6481 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6541 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6601 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
