LOCUS       sequence-466388-        5752 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Secreted human VEGF-A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5752)
  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 5752)
  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: EK0479 CMV-TO-VEGF-A(165) (FLP-IN)
FEATURES             Location/Qualifiers
     source          1..5752
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(841..888)
                     /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     1020..1039
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    1211..1435
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1424..1441
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1497..1526)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2218..2242)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2252..2270
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2273..2291
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(2293..2496)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2322..2342)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2497..2876
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3043..3062
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3137..3241
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3242..3310,3311..4102)
                     /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(3460..3479)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4273..4861
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4762..4781
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5015..5032
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5149..5170
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5185..5190,5191..5208,5209..5215)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5223..5239
                     /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     5228..5250
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5247..5263
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5247..5263
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5312..5433
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5323..5342)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5377..5396
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5563..5752,1..833))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(join(5563..5752,1..833))
                     /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
                     "
ORIGIN
        1 accaagctct gatagagttg gtcaagacca atgcggagca tatacgcccg gagccgcggc
       61 gatcctgcaa gctccggatg cctccgctcg aagtagcgcg tctgctgctc catacaagcc
      121 aaccacggcc tccagaagaa gatgttggcg acctcgtatt gggaatcccc gaacatcgcc
      181 tcgctccagt caatgaccgc tgttatgcgg ccattgtccg tcaggacatt gttggagccg
      241 aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct cggcccaaag catcagctca
      301 tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac
      361 acatggggat cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg accgattcct
      421 tgcggtccga atgggccgaa cccgctcgtc tggctaagat cggccgcagc gatcgcatcc
      481 atggcctccg cgaccggctg cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac
      541 gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa ttccccaatg
      601 tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt gccgataaac ataacgatct
      661 ttgtagaaac catcggcgca gctatttacc cgcaggacat atccacgccc tcctacatcg
      721 aagctgaaag cacgagattc ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg
      781 aacttttcga tcagaaactt ctcgacagac gtcgcggtga gttcaggctt tttggccaag
      841 gaagttccta tactttctag agaataggaa cttcggaata ggaacttcta ggtacgtgaa
      901 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      961 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     1021 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     1081 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtgggg atacccccta
     1141 gagccccagc tggttctttc cgcctcagaa gccatagagc ccaccgcatc cccagaacat
     1201 cgatagatct ccatagagcc caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc
     1261 tcccccttgc tgtcctgccc caccccaccc cccagaatag aatgacacct actcagacaa
     1321 tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc
     1381 aaggaaggca cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgag
     1441 gctgatcagc gggtttaaac tcaatgatga ttatgatgat ggtcgacggc gctattcaga
     1501 tcctcttctg agatgagttt ttgttcgggc cctcctcgag cggctcaccg cctcggctcg
     1561 tcacatctgc aagtacgttc gtttaactca agctgcctcg ccttgcaacg cgagtctgtg
     1621 tttttgcagg aacatttaca cgtctgcgga tcttgtacaa acaaatgctt tctccgctct
     1681 gagcaaggcc cacagggatt ttcttgtctt gctctatctt tctttggtct gcattcacat
     1741 ttgttgtgct gtaggaagct catctctcct atgtgctggc cttggtgagg tttgatccgc
     1801 ataatctgca tggtgatgtt ggactcctca gtgggcacac actccaggcc ctcgtcattg
     1861 cagcagcccc cgcatcgcat caggggcaca caggatggct tgaagatgta ctcgatctca
     1921 tcagggtact cctggaagat gtccaccagg gtctcgattg gatggcagta gctgcgctga
     1981 tagacatcca tgaacttcac cacttcgtga tgattctgcc ctcctccttc tgccatgggt
     2041 gcagcctggg accacttggc atggtggagg tagagcagca aggcaaggct ccaatgcacc
     2101 caagacagca gaaagttcat ggtggcgaag cttaagttta aacgctagag tccggaggct
     2161 ggatcggtcc cggtgtcttc tatggaggtc aaaacagcgt ggatggcgtc tccaggcgat
     2221 ctgacggttc actaaacgag ctcgtcgacg atctctatca ctgataggga gatctctatc
     2281 actgataggg agagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg
     2341 cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa
     2401 caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta
     2461 tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac
     2521 gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc
     2581 gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg
     2641 tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc
     2701 tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg
     2761 cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct
     2821 atgaactaat gaccccgtaa ttgattacta ttaataacta gtcaataatc aatgtcaacg
     2881 cgtatatctg gcccgtacga agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc
     2941 aattgtcggt caagccttgc cttgttgtag cttaaatttt gctcgcgcac tactcagcga
     3001 cctccaacac acaagcaggg agcagatact ggcttaacta tgcggcatca gagcagattg
     3061 tactgagagt gcaccatagg ggatcgggag atctcccgat ccgtcgacgt caggtggcac
     3121 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3181 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3241 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     3301 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     3361 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3421 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3481 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     3541 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     3601 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3661 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3721 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3781 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3841 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3901 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3961 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4021 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     4081 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     4141 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4201 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4261 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4321 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4381 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4441 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4501 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4561 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4621 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4681 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4741 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4801 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4861 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4921 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     4981 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     5041 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     5101 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     5161 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     5221 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
     5281 ctagctagag gtcgacggta tacagacatg ataagataca ttgatgagtt tggacaaacc
     5341 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     5401 tttgtaacca ttataagctg caataaacaa gttggggtgg gcgaagaact ccagcatgag
     5461 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
     5521 ttcatagaag gcggcggtgg aatcgaaatc tcgtagtacg tgctattcct ttgccctcgg
     5581 acgagtgctg gggcgtcggt ttccactatc ggcgagtact tctacacagc catcggtcca
     5641 gacggccgcg cttctgcggg cgatttgtgt acgcccgaca gtcccggctc cggatcggac
     5701 gattgcgtcg catcgaccct gcgcccaagc tgcatcatcg aaattgccgt ca
//
