LOCUS       sequence-459378-        6435 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Membrane-bound mouse renin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6435)
  AUTHORS   Aldrete CA, Call CC, Sant'Anna LE, Vlahos AE, Pei J, Cong Q, Gao XJ
  TITLE     Orthogonalized human protease control of secreted signals.
  JOURNAL   Nat Chem Biol. 2025 Jan 15. doi: 10.1038/s41589-024-01831-x.
  PUBMED    39814991
REFERENCE   2  (bases 1 to 6435)
  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 Jan 15. doi: 10.1038/s41589-024-01831-x."
COMMENT     Sequence Label: CA-0113 CMVTO-Renin(mouse)-ECL
FEATURES             Location/Qualifiers
     source          1..6435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     265..284
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(503..607)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(682..701)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        868..1247
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1248..1451
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1402..1422
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    1453..1471
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1474..1492
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1502..1526
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1618..1627
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2901..2930
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2986..3003)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2992..3216
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3388..3407)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3539..3586
                     /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             3594..4616
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3594..4616
                     /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    4746..4867
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4783..4802)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4837..4856
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4916..4932)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4916..4932)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4929..4951)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4940..4956
                     /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(4964..4970,4971..4988,4989..4994))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5009..5030
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5147..5164)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5318..5906)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5398..5417)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6077..6435,1..433,434..502))
                     /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"
ORIGIN
        1 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
       61 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
      121 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
      181 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
      241 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
      301 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
      361 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
      421 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
      481 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
      541 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
      601 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
      661 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
      721 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
      781 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
      841 gcttcgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
      901 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      961 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     1021 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     1081 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     1141 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1201 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1261 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1321 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1381 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1441 taagcagagc tctccctatc agtgatagag atctccctat cagtgataga gatcgtcgac
     1501 gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc
     1561 atagaagaca ccgggaccga tccagcctcc ggactctagc gtttaaactt aagcttcgcc
     1621 accatggatc gccgtcgaat gccactctgg gctctcttgc tgttatggag tccctgtacc
     1681 ttttccgcta gcccagtggt gctaactaat tacttgaata cacagtatta cggcgagatt
     1741 ggcatcggaa cgccaccgca aacgttcaag gtcatatttg atacagggag tgctaactta
     1801 tgggtgccta gcactaaatg cagtaggctg tacctggcct gcggcatcca ctcactctac
     1861 gaatctagcg actcgtctag ttacatggag aacggatctg actttacaat ccattacgga
     1921 agtggtagag taaaagggtt cttgtcccag gatagtgtga ccgttggcgg cattacagtg
     1981 acccagacct ttggggaggt cacagagctg cctctaattc catttatgct cgcaaaattc
     2041 gacggggtgc tgggcatggg ctttccagca caagccgtgg ggggtgtaac tcctgtgttt
     2101 gaccacatcc tgtctcaggg tgtgctgaag gaagaagtct tctcagtgta ttacaacagg
     2161 ggtagccatt tactgggggg tgaggttgtg ttgggaggtt ccgaccccca gcattatcag
     2221 gggaatttcc actacgtgtc aatttcaaag acagattcct ggcagatcac aatgaagggg
     2281 gtgtcggtgg gcagcagcac cctgctgtgc gaagaagggt gtgccgtagt tgtcgacacc
     2341 ggctctagct tcatttctgc cccgacaagc tccctgaagc tcatcatgca ggcactggga
     2401 gctaaagaga agagaataga ggaatatgtg gttaattgct cccaagtccc caccctgcca
     2461 gatatatctt tcgacctagg aggcagagcc tacactctgt ccagcactga ttatgtcctg
     2521 cagtatccta ataggcggga taaactctgt acgctcgctc tgcacgccat ggatattcct
     2581 ccccctaccg gccccgtctg ggttctcggc gctaccttta tccgcaagtt ctatacagaa
     2641 ttcgaccgac acaacaaccg gatcggattc gccttggcgc ggggcgccgg gacaggaggg
     2701 tctgggggtg gcactggcgg atctggagga gactacaagg acgatgacga caacggcgga
     2761 tcgggggggt ctggtggtag tggcggttcg gggggaggca caggtggatc cctggtcgtg
     2821 gttgctggag tcctgttttg ttatggcttg ctagtgacag tggctctttg tgttgaaaac
     2881 gggggcggct cttagggccc gaacaaaaac tcatctcaga agaggatctg aatagcgccg
     2941 tcgaccatca tcataatcat cattgagttt aaacccgctg atcagcctcg actgtgcctt
     3001 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3061 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3121 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3181 atagcaggca tgctggggat gcggtgggct ctatggagat ctatcgatgt tctggggatg
     3241 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     3301 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3361 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3421 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3481 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtacctaga
     3541 agttcctatt ccgaagttcc tattctctag aaagtatagg aacttccttg gccaaaaagc
     3601 ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg
     3661 acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc
     3721 gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt
     3781 atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca
     3841 gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc
     3901 ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg
     3961 cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat
     4021 acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa
     4081 ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt
     4141 gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg
     4201 tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg
     4261 attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc
     4321 agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg
     4381 cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg
     4441 atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg
     4501 tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag
     4561 tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagcacg
     4621 tactacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4681 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4741 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4801 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4861 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     4921 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4981 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5041 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5101 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5161 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5221 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5281 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5341 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5401 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5461 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5521 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5581 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5641 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5701 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5761 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5821 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5881 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5941 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6001 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6061 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6121 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6181 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6241 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6301 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6361 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6421 ttggtatggc ttcat
//
