LOCUS       sequence-459392-        6453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Membrane-bound human renin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6453)
  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 6453)
  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-0030 CMVTO-Renin-ECL
FEATURES             Location/Qualifiers
     source          1..6453
                     /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"
     CDS             2919..2948
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3004..3021)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3010..3234
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3406..3425)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3557..3604
                     /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             3612..4634
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3612..4634
                     /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    4764..4885
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4801..4820)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4855..4874
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4934..4950)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4934..4950)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4947..4969)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4958..4974
                     /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(4982..4988,4989..5006,5007..5012))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5027..5048
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5165..5182)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5336..5924)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5416..5435)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6095..6453,1..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 accatgcaca gctcagcact gctctgttgc ctggtcctcc tgactggggt gagggctagc
     1681 ctcactctcg gtaatactac ttcctcagtc atcttaacca actacatgga tacgcagtat
     1741 tacggggaaa tcggcattgg cacaccccct cagactttta aggtagtttt tgatactggg
     1801 tctagcaatg tttgggtccc tagcagtaag tgttcccgcc tgtatacagc gtgtgtttac
     1861 cataagttat ttgacgcctc agactcctct agctataaac acaacggcac cgagctcacc
     1921 ctaagataca gcacagggac tgtttcgggg tttctgtccc aggacattat tacagtggga
     1981 ggtattacag tgacccaaat gtttggggag gtgaccgaga tgccggctct gccgttcatg
     2041 ctggcagagt tcgacggggt cgttggcatg ggcttcatcg agcaggctat tgggcgggta
     2101 acccccatct tcgacaacat cattagccag ggcgtgctaa aggaggacgt gttcagcttt
     2161 tattacaata gagattctga aaactctcag agtttgggcg gccagatagt gcttggaggc
     2221 agtgatcccc agcactacga aggcaatttt cactatataa atctgatcaa aactggggtg
     2281 tggcagatcc agatgaaggg ggtgtcagtg ggttcctcta cgttattgtg cgaggacgga
     2341 tgtctggccc tcgtagacac tggagctagt tacatcagcg gctcaaccag ctcaattgaa
     2401 aagctgatgg aggcactggg agcaaaaaag cggttgttcg attacgtcgt gaaatgtaat
     2461 gagggaccta ccctgcccga catctctttc catctgggtg gaaaagagta tactctgaca
     2521 agcgccgatt acgtatttca ggagtcctac tcgtccaaaa aactctgcac gttagcaatc
     2581 cacgccatgg acatccctcc accaaccgga cctacctggg ctctgggggc cacatttata
     2641 cgaaaattct ataccgagtt cgatcgcagg aacaaccgta tcggctttgc acttgccagg
     2701 ggcgccggga caggagggtc tgggggtggc actggcggat ctggaggaga ctacaaggac
     2761 gatgacgaca acggcggatc gggggggtct ggtggtagtg gcggttcggg gggaggcaca
     2821 ggtggatccc tggtcgtggt tgctggagtc ctgttttgtt atggcttgct agtgacagtg
     2881 gctctttgtg ttgaaaacgg gggcggctct tagggcccga acaaaaactc atctcagaag
     2941 aggatctgaa tagcgccgtc gaccatcatc ataatcatca ttgagtttaa acccgctgat
     3001 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3061 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3121 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3181 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggagatct
     3241 atcgatgttc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     3301 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3361 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3421 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3481 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3541 atggttcacg tacctagaag ttcctattcc gaagttccta ttctctagaa agtataggaa
     3601 cttccttggc caaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa
     3661 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca
     3721 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
     3781 acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc
     3841 ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg
     3901 tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg
     3961 ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac
     4021 cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc
     4081 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc
     4141 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg
     4201 atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga
     4261 gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt
     4321 ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag
     4381 gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct
     4441 tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc
     4501 gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga
     4561 ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga
     4621 gggcaaagga atagcacgta ctacgagatt tcgattccac cgccgccttc tatgaaaggt
     4681 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4741 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4801 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4861 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     4921 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4981 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5041 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5101 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5161 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5221 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5281 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5341 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5401 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5461 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5521 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5581 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5641 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5701 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5761 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5821 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5881 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5941 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6001 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6061 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6121 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6181 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6241 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6301 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6361 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6421 tggtgtcacg ctcgtcgttt ggtatggctt cat
//
