LOCUS       sequence-355697-        5796 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HCVP-repressible ER-associated uncaged-nTVMVP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5796)
  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 5796)
  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-p450-nTVMVp-hcvs-AP4
FEATURES             Location/Qualifiers
     source          1..5796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(522..1313,1314..1382))
                     /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     1145..1164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1383..1487)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1562..1581)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1748..2127
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2128..2331
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2282..2302
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2333..2351
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2354..2372
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2382..2406
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2498..2507
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3142..3171
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3227..3244)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3233..3457
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3629..3648)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3780..3827
                     /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             3835..4857
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3835..4857
                     /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    4987..5108
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5024..5043)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5078..5097
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5157..5173)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5157..5173)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5170..5192)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5181..5197
                     /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(5205..5211,5212..5229,5230..5235))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5250..5271
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5388..5405)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5559..5796,1..351))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5639..5658)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
       61 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      121 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      181 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      241 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      301 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      361 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      421 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      481 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      541 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
      601 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
      661 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
      721 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
      781 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
      841 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
      901 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
      961 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1021 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1081 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1141 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1201 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1261 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1321 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1381 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1441 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     1501 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
     1561 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     1621 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     1681 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgtacg ggccagatat
     1741 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     1801 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     1861 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     1921 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     1981 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     2041 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     2101 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     2161 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     2221 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     2281 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctccctatc
     2341 agtgatagag atctccctat cagtgataga gatcgtcgac gagctcgttt agtgaaccgt
     2401 cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga
     2461 tccagcctcc ggactctagc gtttaaactt aagcttcgcc accatggacc ccgttgtcgt
     2521 actgggactc tgcttgtcct gcctgctcct ccttagtttg tggaaacaat cttatggggg
     2581 tggaaaactt ggctccgggg gaggatcctc aaaagcactc ctgaagggag tccgcgactt
     2641 taatcctata tcagcgtgtg tatgtttgct tgaaaactcc tctgatgggc attcagaaag
     2701 gctttttggt attgggtttg gtccctacat aatagcaaac caacacctgt ttcgacgcaa
     2761 taacggcgag cttacgatta agacgatgca cggggaattt aaagtgaaaa acagcacaca
     2821 attgcaaatg aaacccgtcg aagggagaga catcattgta ataaagatgg caaaggattt
     2881 cccacccttt ccgcagaaat tgaaattccg gcaacctact ataaaggatc gagtttgtat
     2941 ggtttctacg aactttcagc aaggaggtgg aggcagtgag gatgtcgtct gctgctcaat
     3001 gtcctacggc agtggaggta cctcacccga ggacgagctt gctgctaatg aagaggagtt
     3061 gcagcaaaat gaacaaaagt tggctcaaat taagcaaaaa cttcaagcta tcaaatacgg
     3121 tagctagctc gaggagggcc cgaacaaaaa ctcatctcag aagaggatct gaatagcgcc
     3181 gtcgaccatc atcataatca tcattgagtt taaacccgct gatcagcctc gactgtgcct
     3241 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3301 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3361 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3421 aatagcaggc atgctgggga tgcggtgggc tctatggaga tctatcgatg ttctggggat
     3481 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3541 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3601 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3661 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3721 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtacctag
     3781 aagttcctat tccgaagttc ctattctcta gaaagtatag gaacttcctt ggccaaaaag
     3841 cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc
     3901 gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg
     3961 cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt
     4021 tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc
     4081 agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg
     4141 cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct
     4201 gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa
     4261 tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa
     4321 actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt
     4381 tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat
     4441 gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg
     4501 gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag
     4561 cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg
     4621 gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc
     4681 gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact
     4741 gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa
     4801 gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac
     4861 gtactacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4921 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4981 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5041 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5101 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     5161 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5221 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5281 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5341 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5401 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5461 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5521 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5581 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5641 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5701 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5761 cgctgtaggt atctcagttc ggtgtaggtc gttcgc
//
