LOCUS       sequence-353271-        6460 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses myc-dL5(E52K, L91S) low affinity Fluorogen Activating 
            Protein (FAP) on the extracellular surface anchored to the mammalian
            cell plasma membrane with the PDGFR transmembrane domain. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6460)
  AUTHORS   Carpenter MA, Wang Y, Telmer CA, Schmidt BF, Yang Z, Bruchez MP
  TITLE     Protein Proximity Observed Using Fluorogen Activating Protein and 
            Dye Activated by Proximal Anchoring (FAP-DAPA) System.
  JOURNAL   ACS Chem Biol. 2020 Aug 17. doi: 10.1021/acschembio.0c00419.
  PUBMED    32786268
REFERENCE   2  (bases 1 to 6460)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Chem 
            Biol. 2020 Aug 17. doi: 10.1021/acschembio.0c00419."
COMMENT     Sequence Label: pcDNA3.1-kappa-myc-dK-2xG4S-TMst
FEATURES             Location/Qualifiers
     source          1..6460
                     /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"
     sig_peptide     949..1011
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     CDS             1018..1047
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1063..1782
                     /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="QAVVTQEPSVTVSPGGTVILTCGSSTGAVTSGHYANWFQQKPGQA
                     PRALIFKTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSLSDVDGYLFGGGT
                     QLTVLSGGGGSGGGGSGGGGSGGGGSQAVVTQEPSVTVSPGGTVILTCGSSTGAVTSGH
                     YANWFQQKPGQAPRALIFKTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSL
                     SDVDGYLFGGGTQLTVLS"
     CDS             1852..1998
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(2051..2068)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2057..2281
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2327..2755
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2414..2433)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2624..2645
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2764..2784)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2769..3098
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2949..3084
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3011..3030
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3165..3959
                     /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(3219..3238)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3829..3848
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4133..4254
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4170..4189)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4224..4243
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4303..4319)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4303..4319)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4316..4338)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4327..4343
                     /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(4351..4357,4358..4375,4376..4381))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4396..4417
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4534..4551)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4705..5293)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4785..4804)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5464..6255,6256..6324))
                     /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     6087..6106
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6325..6429)
                     /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 ttaagcttgc caccggatcc ggcttgggga tatccaccat ggagacagac
      961 acactcctgc tatgggtact gctgctctgg gttccaggtt ccactggtga cgaattcgag
     1021 cagaagctga tcagcgagga ggacctgggc ctttctggcc tgcaggccgt cgttacccaa
     1081 gaacctagtg ttaccgttag cccaggtggt actgtcatac ttacttgtgg aagttctacg
     1141 ggtgccgtca catctggtca ttatgcaaat tggtttcaac aaaaaccagg acaagctcca
     1201 agagctttga tttttaaaac tgataagaag tattcttgga ccccaggtag attttctgga
     1261 tctttgctgg gagcaaaggc agctttgaca atatcagatg ctcagcctga ggacgaagcc
     1321 gagtattact gttctcttag cgacgtggat ggctacttgt ttggcggtgg aacacaactg
     1381 acggttctgt ccggtggtgg aggatctggt ggaggtggta gcggaggtgg aggttcagga
     1441 ggcggcggtt ctcaggctgt ggtgactcag gagccgtcag tgactgtgtc cccaggaggg
     1501 acagtcattc tcacttgtgg ctccagcact ggagctgtca ccagcggtca ttatgccaac
     1561 tggttccagc agaagcctgg ccaagccccc agggcactta tatttaaaac cgacaagaaa
     1621 tattcctgga cccctggccg attctcaggc tccctccttg gggccaaggc tgccctgacc
     1681 atctcggatg cgcagcctga agatgaggct gagtattact gttcgctctc cgacgtagac
     1741 ggttatctgt tcggaggagg cacccagctg accgtcctct ccgcggccgg tctggcccat
     1801 atgggaggcg gtgggtctgg tggcggtggc agcaccggta tcttactcga ggctgtgggc
     1861 caggacacgc aggaggtcat cgtggtgcca cactccttgc cctttaaggt ggtggtgatc
     1921 tcagccatcc tggccctggt ggtgctcacc atcatctccc ttatcatcct catcatgctt
     1981 tggcagaaga agccacgtta agtcgaggca gctgtcgact ctagagggcc cgtttaaacc
     2041 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2101 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2161 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2221 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2281 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     2341 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2401 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2461 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2521 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2581 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2641 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2701 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     2761 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2821 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2881 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     2941 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3001 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3061 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3121 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3181 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3241 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3301 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3361 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3421 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3481 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3541 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3601 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3661 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     3721 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     3781 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     3841 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     3901 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     3961 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4021 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4081 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4141 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4201 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4261 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     4321 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4381 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4441 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4501 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4561 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4621 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4681 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4741 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4801 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4861 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4921 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4981 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5041 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5101 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5161 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5221 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5281 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5341 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5401 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5461 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5521 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5581 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5641 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5701 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5761 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5821 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5881 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5941 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6001 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6061 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6121 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6181 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6241 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6301 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6361 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6421 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
