LOCUS       sequence-492999-        7373 bp    DNA     circular SYN 18-NOV-2025
DEFINITION  Expresses MTS-FAP-GFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7373)
  AUTHORS   Moore WM, Brea RJ, Knittel C, Wrightsman E, Hui B, Lou J, Ancajas 
            CF, Best MD, Obara, CJ Devaraj NK, Budin I
  TITLE     Leaflet-specific phospholipid imaging using genetically encoded 
            proximity sensors
  JOURNAL   Nat Chem Biol (2025). https://doi.org/10.1038/s41589-025-02021-z.
  PUBMED    38746395
REFERENCE   2  (bases 1 to 7373)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 18, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol (2025). https://doi.org/10.1038/s41589-025-02021-z."
COMMENT     Sequence Label: pSBbi-MTS-FAP-GFP
FEATURES             Location/Qualifiers
     source          1..7373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     76..95
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(195..794)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(278..298)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     775..794
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        1426..2608
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     2556..2576
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(3654..3656,3657..3659,3660..4370)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3654..3656,3657..3659,3660..4370)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3699..3720)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3960..3979)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4307..4328
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4399..4416)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4405..4629
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        5061..5165
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5166..5234,5235..6026)
                     /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     complement(5384..5403)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6197..6785
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6686..6705
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6939..6956
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7330..7349
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    join(7348..7373,1..30)
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
ORIGIN
        1 ctattgcaat gaaaataaat ttcctttatt agccagaagt cagatgctca aggggcttca
       61 tgatgtcccc ataatttttg gcagagggaa aaagatctca gtggtatttg tgagccaggg
      121 cattggccac accagccacc accttctgat aggcagcctg cacctgagga gtgaattcac
      181 gacaggcctt cgaatcaggc accgggcttg cgggtcatgc accaggtgcg cggtccttcg
      241 ggcacctcga cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg
      301 ggcgcggagg tctccaggaa ggcgggcacc ccggcgcgct cggccgcctc cactccgggg
      361 agcacgacgg cgctgcccag acccttgccc tggtggtcgg gcgagacgcc gacggtggcc
      421 aggaaccacg cgggctcctt gggccggtgc ggcgccagga ggccttccat ctgttgctgc
      481 gcggccagcc gggaaccgct caactcggcc atgcgcgggc cgatctcggc gaacaccgcc
      541 cccgcttcga cgctctccgg cgtggtccag accgccaccg cggcgccgtc gtccgcgacc
      601 cacaccttgc cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc
      661 cgctcgatgt ggcggtccgg atcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg
      721 gcggcgaggg tgcgtacggc cctggggacg tcgtcgcggg tggcgaggcg caccgtgggc
      781 ttgtactcgg tcatggtggc ctcaggtgca gaggtttcta cagggaaaga gagaaaggct
      841 acagttagtt tgctgtgatg atgctaagtg ccgaggtcag atgtctatgg attgtacacg
      901 gaatcccaac gcaggaacca aaaaacactc acccaaaaaa atggcgctaa ggccgagaga
      961 aaggcgaagt tccgtctacg gctatatacc ttgaaacagg ggcggcagaa gtgacgcaac
     1021 gcggttgcgc ggacggcttc ctgtgttttc attggtcgaa gcgttctgcg cgtgcgcaga
     1081 gattccccag ggaaggcggg ccctcttccg aatgtccaat ccggacattc ttccggttgg
     1141 acctccgcgg tcagcgctgt gcccggcgag agatcacgtg gggcgcggag gcggtgctgc
     1201 tggggcacgg ccgtccagcc tcggcggcca tatttttgag gggctgttca tctcgttcac
     1261 acgctctgtc cgccatgttt gtgagtggaa gcgccattac cccttcaagc gactgaaggc
     1321 tgcagggcct ctggtggccc gcatggggag accagacccg ccaggcccgc ctttccgcac
     1381 tcagtccggg cttactttat tttgtgagac agggtctcgc ctagaggctc cggtgcccgt
     1441 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     1501 tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     1561 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     1621 gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc
     1681 gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc cacgcccctg
     1741 gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag
     1801 gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ttgggcgctg
     1861 gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc
     1921 tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct
     1981 tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga
     2041 cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc
     2101 gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc
     2161 gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc
     2221 ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc
     2281 gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt
     2341 cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag
     2401 cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca
     2461 tactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga
     2521 atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag
     2581 tttttttctt ccatttaagg tgtcgtgaaa actaccccaa gctggcctct gaggccacca
     2641 tgctgtctaa tttgagaatc ctgctcaaca aggcagctct tagaaaggct cacacttcca
     2701 tggttcgaaa ttttcggtat gggaagccag tccaggaatt cggcctttct ggcctgcaag
     2761 tccagttgca agaatctgga cctggtttgg ttaaaccatc tgagacactt tccctaacct
     2821 gcacagtcag tggcgcatca atttcctcct cgcactatta ttggggttgg attagacaac
     2881 cacctggaaa gggcccagag tggatagggt ctatgtacta ctcgggtaga acttactata
     2941 atcctgcctt gaagtcacgt gttaccatat cctctgataa gtcaaaaaac caattctttt
     3001 taaaactcac tagcgttacc gctgctgata cagctgtcta ttatgctgcc cgtgagggcc
     3061 caacgcatta ttatgataat tcaggcccaa tacctagcga tgagtatttt caacattggg
     3121 gccagggcac tttagttaca gtttccggtg gtggcggctc tggtggcggt ggcagcggcg
     3181 gtggtggttc cggaggcggc ggttctcagg tgcagctgca ggagtcgggc ccaggactgg
     3241 tgaagccttc ggagaccctg tccctcacct gcactgtctc tggtgcctcc atcagcagta
     3301 gtcattacta ctggggctgg attcgccagc ccccagggaa ggggcctgag tggattggga
     3361 gtatgtatta tagtgggaga acgtactaca acccggccct caagagtcga gtcaccatat
     3421 catcagacaa gtcgaagaac cagttcttct tgaagttgac ctctgtaacc gccgcggaca
     3481 cggccgtgta ttacgctgcg agggagggac ccacacatta ctatgataat agtggtccaa
     3541 taccttcgga tgagtatttc cagcactggg gccagggtac cctggtcact gtctccgcgg
     3601 ccggtctggc ccatatggga ggcggtgggt ctggtggcgg tggcagcacc ggtatggtga
     3661 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     3721 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     3781 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     3841 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     3901 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     3961 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     4021 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     4081 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     4141 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     4201 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     4261 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     4321 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag ctcgagtaat
     4381 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     4441 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     4501 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     4561 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     4621 gctctatggc ttctgaggcg gaaagaacca gctgggccta ctagatagcg gaccccttac
     4681 cgaaacatcg ccgcattctg cagaggagtc gaacttagtg tatgtaaact tctgacccac
     4741 tggaattgtg atacagtgaa ttataagtga aataatctgt ctgtaaacaa ttgttggaaa
     4801 aatgacttgt gtcatgcaca aagtagatgt cctaactgac ttgccaaaac tattgtttgt
     4861 taacaagaaa tttgtggagt agttgaaaaa cgagttttaa tgactccaac ttaagtgtat
     4921 gtaaacttcc gacttcaact gtatagggat ccaattttaa caaaatatta acgcttacaa
     4981 tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatcaggtg
     5041 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5101 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5161 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5221 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5281 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5341 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5401 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5461 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5521 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5581 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5641 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5701 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5761 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5821 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5881 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5941 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6001 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6061 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6121 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6181 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6241 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6301 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6361 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6421 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6481 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6541 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6601 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6661 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6721 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6781 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6841 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6901 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     6961 cggatcccta tacagttgaa gtcggaagtt tacatacacc ttagccaaat acatttaaac
     7021 tcactttttc acaattcctg acatttaatc ctagtaaaaa ttccctgtct taggtcagtt
     7081 aggatcacca ctttatttta agaatgtgaa atatcagaat aatagtagag agaatgattc
     7141 atttcagctt ttatttcttt catcacattc ccagtgggtc agaagtttac atacactcga
     7201 ctcctctgca gaatgcggcg atgtttcggt aaggggtccc gagtagctag ttcatggcag
     7261 ccagcatatg gcatatgttg ccaaactcta aaccaaatac tcattctgat gttttaaatg
     7321 atttgccctc ccatatgtcc ttccgagtga gagacacaaa aaattccaac aca
//
