LOCUS       sequence-466379-        8765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optimized VEGF-sensing eLIDAR with VEGFR2 ecto+TM domains fused with
            tdMCP, with Golgi traffic sequence and ER export sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8765)
  AUTHORS   Zhang X, Mille-Fragoso LS, Kaseniit KE, Lee AP, Zhang M, Call CC, Hu
            Y, Xie Y, Gao XJ
  TITLE     Post-transcriptional modular synthetic receptors.
  JOURNAL   Nat Chem Biol. 2025 Mar 28. doi: 10.1038/s41589-025-01872-w.
  PUBMED    40155716
REFERENCE   2  (bases 1 to 8765)
  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"; journalName: "Nat Chem 
            Biol. 2025 Mar 28. doi: 10.1038/s41589-025-01872-w."
COMMENT     Sequence Label: XZ362
            EF1a-VEGFR2sp-HA-VEGFR2(Ecto+TM)-TS-tdMC...
FEATURES             Location/Qualifiers
     source          1..8765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2974..3000)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      3063..3072
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          3094..4032
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(3117..3137)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        complement(4033..4244)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     primer_bind     4371..4390
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4465..4569
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4570..4638,4639..5430)
                     /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(4788..4807)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5601..6189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6090..6109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6343..6360
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6477..6498
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6513..6518,6519..6536,6537..6543)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6551..6567
                     /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)."
     primer_bind     6556..6578
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6575..6591
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6575..6591
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6640..6761
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6651..6670)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6705..6724
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6891..7913)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(6891..7913)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(7921..7968)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     8100..8119
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    8291..8515
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     8504..8521
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(8577..8606)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 caattccata ttcaagtatg agcgccatgc acctttgggg acctccactt taatggtata
       61 cttccggttc tgggcactag attggcgtac actacaagtc accttatagg cttgtgatct
      121 tgaattggaa gagatccact cggctatccc attcgcaaag ttacttggag ctactgtgac
      181 gtctccagtg ccgccgttgt ccacgagcac aaactgtgta aaattggagg ccatggaccc
      241 gccaccggtt ccgccaccat atattcctga gttggcggcg atcgcagagg gaatcgggtt
      301 accgtctttc aaaagacctt gcatggcctt gacaatcagc tcgcagtctg aattagtcgc
      361 aaagattggg atcgtaagct ccatattaag ataagatcgc cacgcccctt ttggcacttc
      421 tacctttatg gtatattttc ggttctgtgc ggaactttgg cgaactgagc atgtgacttt
      481 atatgcctga ctccggctgt tactacttat ccattctgca atcccgttcg caaaattgct
      541 tggcgcaaca gttacgtctc cggtgccacc attgtcaacc aacacaaatt gcgtaaagtt
      601 gctcgccatc acgttgatgt cgatctggtc cagggggatg tactcgccct cgctggtgat
      661 cctgctcttg gcggccgcgg taccgatgat gacaagaagt agccagaaga acatggcaat
      721 caccgccgtg cctactagaa taatgatttc caagttcgtc ttttcctggg caccttctat
      781 tatgaaaaat gcctccactt ttgcacagcc aagaacactg catgcctggc aggtgtagag
      841 gccttcgtcc tccttcctca ctctgcggat agtgaggttc cggttcccat ccttcaatac
      901 aatgcctgag tcttctacaa gggtctcatt atctttaaac cacatgatct gtggaggggg
      961 attcccagat gccgtgcatg agacttcgat gctttcccca atacttgtcg tctgattctc
     1021 caggtttcct gtgatcgtgg gtgccacacg ctctaggact gtgagctgcc tgaccacgca
     1081 atgtcttttc ttggtcttcc tgtcttgagc aaggcagaca tagtctcctt ggtcctgcaa
     1141 ggatgcattc ttaagctcca tgatcaaaat gtcatttgtg ctattagaga acatggtggc
     1201 attcaatttc caaagagtat ccaagttctt gcaaacaggt gtgggcaact ctcccacatg
     1261 gattggcaga ggctgtgggc caagcttgta ccatgtgagg ttctcaaacg tagatctgtc
     1321 tgcagtgcac cacaaagaca cgctctcctg ctcagtgggc tgcatgtcag gttgcaaagt
     1381 aatttcagga cccctggtca cgtggaagga gatcaccctc tctcctctcc cgactttgtt
     1441 gaccgcttca catttgtaca aagctgacac atttgccgct tggataacaa gggtacttac
     1501 agttttgttt tttccttcaa ttagagcaaa ttgattttta ttaacttcaa ttttatttcc
     1561 tccctggaag tcctccacac ttctccattc ttcacaaggg tatgggtttg tcactgagac
     1621 agcttggctg ggctcgttgg cgcactcttc ctccaactgc caataccagt ggatgtgatg
     1681 cgggggagga atggcataga ccgtacatgt cagcgtttga gtggtgccgt actggtagga
     1741 atccacagga gagattagag atttctcacc aatctggggt gggacataca caaccagaga
     1801 gaccacatgg ctctgcttct cctttgaaat gggattggta aggatgacag tgtaatttcc
     1861 tgtgtctctt tcactcactt ccataatcgt cagtacatgc cccgctttaa ttgtgtgatt
     1921 ggactcaagg ggtattccat ttttatacca ttttatttct gggggtgggt aaccaaggta
     1981 cttcgcaggg attctgacac gctcccccac cgtggcttcc accagagatt ccatgccact
     2041 tccaaaagca acaaaaggtt tttcatggac cctgacaaat gtgctgttct tcttggtcat
     2101 cagcccactg gatgctgcac aggtgtacaa tccttggtca ctccgggtta caccatctat
     2161 agttaaggtg ctcaaaaatt tcttcatctc actcccagac tgggttttta ggtctcggtt
     2221 tacaagtttc ttatgctgat gcttcgaaga agggtattcc cagttgaagt caatccccac
     2281 atttagttca gttcttgctg tacaatttaa gacaagcttt tctccaacag atagttcaat
     2341 tccatgagac ggactcagaa ccacatcata aatcctatac cctacaacga caactatgta
     2401 cataatagac tggtaacttt catcattaat ttttgcttca cagaagacca tgccagcata
     2461 gctgatcatg tagctgggaa tagtaaagcc cttcttgctg tcccaggaaa ttctgttacc
     2521 atcaggaaca aatctctttt ctgggtatct tgcacaaagt gacacgttga gatttgaaat
     2581 ggacccgaga catggaatca ccacagtttt gtttttgttc tcagtaatgt acacgactcc
     2641 atgttggtca ctaacagaag caataaatgg agatctgtaa tcttgaacat agacataaat
     2701 gaccgaggcc aagtcagttt cccggtagaa gcacttgtag gctccagtgt catttccgat
     2761 cacttttgga attgtgagtg tcttacagaa gaggccatcg ctgcactcag tcacctccac
     2821 cctttgctca ctgccactct gattattggg ccaaagccag tccaagtccc tctgtcccct
     2881 gcaagtaatt tgaagagttg tattagcctt aattgtaagt atgtcttttt gtatgctgag
     2941 cctgggcaga tcaagagaaa cactaggcaa accagcgtaa tctggaacat cgtatgggta
     3001 cacagaggcg gcccgggtct ccacgcagag ccacagggcg acggccagca gcaccttgct
     3061 ctccatggtg gcgaactagt agcttcacga cacctgaaat ggaagaaaaa aactttgaac
     3121 cactgtctga ggcttgagaa tgaaccaaga tccaaactca aaaagggcaa attccaagga
     3181 gaattacatc aagtgccaag ctggcctaac ttcagtctcc acccactcag tgtggggaaa
     3241 ctccatcgca taaaacccct ccccccaacc taaagacgac gtactccaaa agcgcgagaa
     3301 ctaatcgagg tgcctggacg gcgcccggta ctccgtggag tcacatgaag cgacggctga
     3361 ggacggaaag gcccttttcc tttgtgtggg tgactcaccc gcccgctctc ccgagcgccg
     3421 cgtcctccat tttgagctcc ctgcagcagg gccgggaagc ggccatcttt ccgctcacgc
     3481 aactggtgcc gaccgggcca gccttgccgc ccagggcggg gcgatacacg gcggcgcgag
     3541 gccaggcacc agagcaggcc ggccagcttg agactacccc cgtccgattc tcggtggccg
     3601 cgctcgcagg ccccgcctcg ccgaacatgt gcgctgggac gcacgggccc cgtcgccgcc
     3661 cgcggcccca aaaaccgaaa taccagtgtg cagatcttgg cccgcattta caagactatc
     3721 ttgccagaaa aaaagcgtcg cagcaggtca tcaaaaattt taaatggcta gagacttatc
     3781 gaaagcagcg agacaggcgc gaaggtgcca ccagattcgc acgcggcggc cccagcgccc
     3841 aggccaggcc tcaactcaag cacgaggcga aggggctcct taagcgcaag gcctcgaact
     3901 ctcccaccca cttccaaccc gaagctcggg atcaagaatc acgtactgca gccaggtgga
     3961 agtaattcaa ggcacgcaag ggccataacc cgtaaagagg ccaggcccgc gggaaccaca
     4021 cacggcactt acctgtgttc tggcggcaaa cccgttgcga aaaagaacgt tcacggcgac
     4081 tactgcactt atatacggtt ctcccccacc ctcgggaaaa aggcggagcc agtacacgac
     4141 atcactttcc cagtttaccc cgcgccacct tctctaggca ccggttcaat tgccgacccc
     4201 tccccccaac ttctcgggga ctgtgggcga tgtgcgctct gcccactgac gggcacgatt
     4261 cttcatgcaa ttgtcggtca agccttgcct tgttgtagct taaattttgc tcgcgcacta
     4321 ctcagcgacc tccaacacac aagcagggag cagatactgg cttaactatg cggcatcaga
     4381 gcagattgta ctgagagtgc accatagggg atcgggagat ctcccgatcc gtcgacgtca
     4441 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4501 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4561 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4621 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4681 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4741 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4801 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4861 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4921 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4981 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5041 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5101 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5161 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5221 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5281 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5341 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5401 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5461 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5521 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5581 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5641 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5701 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5761 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5821 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5881 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5941 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6001 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6061 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6121 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6181 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6241 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6301 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6361 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6421 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     6481 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     6541 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     6601 gccaagctct agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg
     6661 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     6721 ttgctttatt tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc
     6781 agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag
     6841 cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtagtacgtg ctattccttt
     6901 gccctcggac gagtgctggg gcgtcggttt ccactatcgg cgagtacttc tacacagcca
     6961 tcggtccaga cggccgcgct tctgcgggcg atttgtgtac gcccgacagt cccggctccg
     7021 gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa attgccgtca
     7081 accaagctct gatagagttg gtcaagacca atgcggagca tatacgcccg gagccgcggc
     7141 gatcctgcaa gctccggatg cctccgctcg aagtagcgcg tctgctgctc catacaagcc
     7201 aaccacggcc tccagaagaa gatgttggcg acctcgtatt gggaatcccc gaacatcgcc
     7261 tcgctccagt caatgaccgc tgttatgcgg ccattgtccg tcaggacatt gttggagccg
     7321 aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct cggcccaaag catcagctca
     7381 tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac
     7441 acatggggat cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg accgattcct
     7501 tgcggtccga atgggccgaa cccgctcgtc tggctaagat cggccgcagc gatcgcatcc
     7561 atggcctccg cgaccggctg cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac
     7621 gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa ttccccaatg
     7681 tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt gccgataaac ataacgatct
     7741 ttgtagaaac catcggcgca gctatttacc cgcaggacat atccacgccc tcctacatcg
     7801 aagctgaaag cacgagattc ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg
     7861 aacttttcga tcagaaactt ctcgacagac gtcgcggtga gttcaggctt tttggccaag
     7921 gaagttccta tactttctag agaataggaa cttcggaata ggaacttcta ggtacgtgaa
     7981 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     8041 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     8101 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     8161 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtgggg atacccccta
     8221 gagccccagc tggttctttc cgcctcagaa gccatagagc ccaccgcatc cccagaacat
     8281 cgatagatct ccatagagcc caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc
     8341 tcccccttgc tgtcctgccc caccccaccc cccagaatag aatgacacct actcagacaa
     8401 tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc
     8461 aaggaaggca cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgag
     8521 gctgatcagc gggtttaaac tcaatgatga ttatgatgat ggtcgacggc gctattcaga
     8581 tcctcttctg agatgagttt ttgttcgggc cctcctcgag cggctcacac ctcgttctcg
     8641 tagcagaagt aaattccgct attcgcagcg attgcggaag gtatcgggtt gccatctttc
     8701 agaagtcctt gcatggcttt tacaataagc tcgcagtcgc tattggtcgc aaatatcgga
     8761 atcgt
//
