LOCUS       sequence-208311-        6675 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 6675)
  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 6675)
  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..6675
                     /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"
     promoter        619..822
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     773..793
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     867..886
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        867..885
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1494..2213
                     /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(2269..2286)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2275..2499
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2545..2973
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2632..2651)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2842..2863
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2982..3002)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2987..3316
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3167..3302
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3229..3248
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3383..4177
                     /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(3437..3456)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4047..4066
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4351..4472
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4388..4407)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4442..4461
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4521..4537)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4521..4537)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4534..4556)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4545..4561
                     /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(4569..4575,4576..4593,4594..4599))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4614..4635
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4752..4769)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4923..5508)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5003..5022)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5679..6470,6471..6539))
                     /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     6302..6321
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6540..6644)
                     /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 atcatatata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      541 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      601 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      661 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      721 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      781 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa
      841 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagctggc
      901 tagcgtttaa acttaagctt gccaccggat cccttgaatt catggactcg ggccgggact
      961 tcctgaccct gcacggccta caggatgatg aggatctaca ggcgctgctg aagggcagcc
     1021 agctcctgaa ggtgaagtcc agctcatgga ggagagagcg gttctacaag ttgcaggagg
     1081 actgcaagac catctggcag gagtcccgca aggtcatgcg gaccccggag tcccagctgt
     1141 tctccatcga ggacattcag gaggtgcgaa tggggcaccg cacggagggt ctggagaagt
     1201 tcgcccgtga tgtgcccgag gaccgctgct tctccattgt cttcaaggac cagcgcaata
     1261 cactagacct catcgcccca tcgccagctg atgcccagca ctgggtgctg gggctgcaca
     1321 agatcatcca ccactcaggc tccatggacc agcgtcagaa gctacagcac tggattcact
     1381 cctgcttgcg aaaagctgac aaaaacaagg acaacaagat gagcttcaag gagctgcaga
     1441 acttcctgaa gcatatggga ggcggtgggt ctggtggcgg tggcagcacc ggtcaggccg
     1501 tcgttaccca agaacctagt gttaccgtta gcccaggtgg tactgttata cttacttgtg
     1561 gaagtggtac gggtgccgtc acatctggtc attatgcaaa ttggtttcaa caaaaaccag
     1621 gacaagctcc aagagctttg atttttgata ctgataagaa gtattcttgg accccaggta
     1681 gattttctgg atctttgctg ggagcaaagg cagctttgac aatatcagat gctcagcctg
     1741 aggacgaagc cgagtattac tgttctctta gcgacgtgga tggctacttg tttggcggtg
     1801 gaacacaact gacggttctg tccggtggtg gaggatctgg tggaggtggt agcggaggtg
     1861 gaggttcagg aggcggcggt tctcaggctg tggtgactca ggagccgtca gtgactgtgt
     1921 ccccaggagg gacagtcatt ctcacttgtg gctccggcac tggagctgtc accagtggtc
     1981 attatgccaa ctggttccag cagaagcctg gccaagcccc cagggcactt atatttgaca
     2041 ccgacaagaa gtattcctgg acccctggcc gattctcagg ctccctcctt ggggccaagg
     2101 ctgccctgac catctcggat gcgcagcctg aagatgaggc tgagtattac tgttcgctct
     2161 ccgacgttga cggttatctg ttcggaggag gcacccagct gaccgtcctc tcctaatagc
     2221 tcgaggcagc tgtcgactct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc
     2281 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     2341 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     2401 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     2461 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     2521 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2581 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2641 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2701 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2761 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2821 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2881 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2941 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3001 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3061 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3121 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3181 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3241 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3301 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     3361 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3421 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3481 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3541 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     3601 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3661 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3721 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3781 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3841 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3901 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3961 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4021 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4081 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4141 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4201 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4261 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4321 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     4381 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4441 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     4501 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4561 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4621 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4681 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4741 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4801 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4861 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4921 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4981 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5041 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5101 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5161 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5221 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5281 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5341 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5401 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5461 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5521 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5581 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5641 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5701 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5761 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5821 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5881 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5941 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6001 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6061 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6121 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6181 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6241 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6301 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6361 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6421 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6481 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6541 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6601 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6661 gtgccacctg acgtc
//
