LOCUS       sequence-355672-        5799 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ras Binding Domain split-cTEVP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5799)
  AUTHORS   Vlahos AE, Kang J, Aldrete CA, Zhu R, Chong LS, Elowitz MB, Gao XJ
  TITLE     Protease-controlled secretion and display of intercellular signals.
  JOURNAL   Nat Commun. 2022 Feb 17;13(1):912. doi: 10.1038/s41467-022-28623-y.
  PUBMED    35177637
REFERENCE   2  (bases 1 to 5799)
  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 
            Commun."; date: "2022-02-17"; volume: "13(1):912. doi"; pages: " 
            10.1038/s41467-022-28623-y"
COMMENT     Sequence Label: CMVTO-RBDK65E-cTEVp
FEATURES             Location/Qualifiers
     source          1..5799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(591..1382,1383..1451))
                     /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     1214..1233
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1452..1556)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1631..1650)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1817..2196
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2197..2400
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2351..2371
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2402..2420
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2423..2441
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2451..2475
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2567..2576
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3214..3243
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3299..3316)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3305..3529
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3701..3720)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3852..3899
                     /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)."
     CDS             3907..4929
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3907..4929
                     /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
                     "
     polyA_signal    5059..5180
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5096..5115)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5150..5169
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5229..5245)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5229..5245)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5242..5264)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5253..5269
                     /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(5277..5283,5284..5301,5302..5307))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5322..5343
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5460..5477)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5631..5799,1..420))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5711..5730)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
       61 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
      121 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
      181 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
      241 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
      301 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
      361 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
      421 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
      481 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      541 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      601 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      661 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      721 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      781 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      841 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      901 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      961 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     1021 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     1081 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     1141 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     1201 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     1261 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     1321 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     1381 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1441 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1501 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1561 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     1621 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     1681 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     1741 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgtacgg
     1801 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
     1861 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     1921 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     1981 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     2041 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     2101 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     2161 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     2221 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     2281 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     2341 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     2401 ctccctatca gtgatagaga tctccctatc agtgatagag atcgtcgacg agctcgttta
     2461 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
     2521 cgggaccgat ccagcctccg gactctagcg tttaaactta agcttcgcca ccatggatcc
     2581 gagtaagacg tcaaacacaa tcagggtttt tctccccaat gaacagcgga ccgtggtaaa
     2641 cgtccgaaac gggatgagtc tccacgactg tctgatgaaa gccttgaagg tgagggggct
     2701 tcagcctgaa tgttgcgctg tctttcgcct tctccacgaa cacaagggcg caaaggcgag
     2761 gcttgactgg aatacagacg ctgcaagttt gattggggaa gaacttcagg tcgatttcct
     2821 caccggtggc tcagggagca agagtatgag ttctatggtg tccgacacta gctgcacgtt
     2881 cccctcaagt gatgggatat tctggaaaca ctggatacag actaaagacg gacagtgtgg
     2941 aagcccattg gtttccaccc gagatggttt tatcgtgggt atccatagcg cctctaattt
     3001 cacaaacacg aacaactatt tcacttcagt gcccaaaaac tttatggagc tgctcacaaa
     3061 ccaagaagcg cagcagtggg taagcggttg gagacttaac gctgactcag ttctctgggg
     3121 ggggcacaaa gtattcatgg taaagccaga ggagccattc caaccagtca aagaagccac
     3181 acaacttatg aacagctagc tcgaggaggg cccgaacaaa aactcatctc agaagaggat
     3241 ctgaatagcg ccgtcgacca tcatcataat catcattgag tttaaacccg ctgatcagcc
     3301 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3361 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3421 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3481 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatgga gatctatcga
     3541 tgttctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3601 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3661 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3721 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3781 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3841 tcacgtacct agaagttcct attccgaagt tcctattctc tagaaagtat aggaacttcc
     3901 ttggccaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
     3961 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
     4021 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
     4081 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac
     4141 attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg
     4201 ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg
     4261 gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa
     4321 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg
     4381 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat
     4441 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc
     4501 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
     4561 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg
     4621 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
     4681 ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
     4741 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
     4801 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat
     4861 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
     4921 aaggaatagc acgtactacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4981 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5041 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5101 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5161 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     5221 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5281 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5341 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5401 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5461 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5521 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5581 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5641 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5701 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5761 ccgaccctgc cgcttaccgg atacctgtcc gcctttctc
//
