LOCUS       sequence-489490-        4765 bp    DNA     circular SYN 16-OCT-2025
DEFINITION  Constitutive expression of human VEGFA-121 isoform.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4765)
  AUTHORS   Edelstein HI, Cosio A, Ezekiel ML, Corcoran WK, Morris AH, Leonard 
            JN
  TITLE     Conversion of natural cytokine receptors into orthogonal synthetic 
            biosensors.
  JOURNAL   Nat Chem Biol. 2025 Aug 22:10.1038/s41589-025-01986-1. doi: 
            10.1038/s41589-025-01986-1.
  PUBMED    40846994
REFERENCE   2  (bases 1 to 4765)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol. 2025 Aug 22:10.1038/s41589-025-01986-1. doi: 
            10.1038/s41589-025-01986-1."
COMMENT     Sequence Label: L4869 hVEGFA-121 in pcDNA backbone
FEATURES             Location/Qualifiers
     source          1..4765
                     /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..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1395..1412)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1401..1625
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1671..2099
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1758..1777)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1968..1989
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2122..2318
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2169..2304
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2231..2250
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2438..2559
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2475..2494)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2529..2548
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2608..2624)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2608..2624)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2621..2643)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2632..2648
                     /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(2656..2662,2663..2680,2681..2686))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2701..2722
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2839..2856)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3010..3598)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3090..3109)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3769..4560,4561..4629))
                     /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     4392..4411
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4630..4734)
                     /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 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagtcccaa gctggctagc
      901 gccaccatga actttctgct gtcttgggtg cattggagcc ttgccttgct gctctacctc
      961 caccatgcca agtggtccca ggctgcaccc atggcagaag gaggagggca gaatcatcac
     1021 gaagtggtga agttcatgga tgtctatcag cgcagctact gccatccaat cgagaccctg
     1081 gtggacatct tccaggagta ccctgatgag atcgagtaca tcttcaagcc atcctgtgtg
     1141 cccctgatgc gatgcggggg ctgctgcaat gacgagggcc tggagtgtgt gcccactgag
     1201 gagtccaaca tcaccatgca gattatgcgg atcaaacctc accaaggcca gcacatagga
     1261 gagatgagct tcctacagca caacaaatgt gaatgcagac caaagaaaga tagagcaaga
     1321 caagaaaaat gtgacaagcc gaggcggtga gcggccgctc gagtctagag ggcccgttta
     1381 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     1441 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     1501 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     1561 ggacagcaag ggggaggatt gggaagtcaa tagcaggcat gctggggatg cggtgggctc
     1621 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     1681 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     1741 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     1801 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     1861 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     1921 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     1981 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2041 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     2101 attctgtgga atgtgtgtca gttaggctca attagtcagc aaccatagtc ccgcccctaa
     2161 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2221 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     2281 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     2341 ccattttcgg aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     2401 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     2461 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2521 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     2581 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     2641 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     2701 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2761 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2821 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2881 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2941 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3001 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3061 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3121 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3181 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3241 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3301 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3361 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3421 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3481 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3541 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3601 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3661 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3721 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3781 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3841 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3901 gataccgcga gatccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3961 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4021 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4081 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4141 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4201 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4261 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4321 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4381 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4441 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4501 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4561 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4621 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4681 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4741 tccccgaaaa gtgccacctg acgtc
//
