LOCUS       sequence-356774-        5649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Uncaged-cTVMVP with P3 leucine zipper.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5649)
  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 5649)
  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-P3-cTVMVP
FEATURES             Location/Qualifiers
     source          1..5649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        53..432
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        433..636
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     587..607
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    638..656
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    659..677
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     687..711
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      803..812
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1300..1329
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1385..1402)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1391..1615
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1787..1806)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    1938..1985
                     /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             1993..3015
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             1993..3015
                     /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    3145..3266
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3182..3201)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3236..3255
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3315..3331)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3315..3331)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3328..3350)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3339..3355
                     /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(3363..3369,3370..3387,3388..3393))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3408..3429
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3546..3563)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3717..4305)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3797..3816)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4476..5267,5268..5336))
                     /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     5099..5118
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5337..5441)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5516..5535)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgcttagggt taggcgtttt gcgctgcttc gtacgggcca gatatacgcg ttgacattga
       61 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      121 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      181 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      241 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      301 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      361 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      421 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      481 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      541 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      601 aggcgtgtac ggtgggaggt ctatataagc agagctctcc ctatcagtga tagagatctc
      661 cctatcagtg atagagatcg tcgacgagct cgtttagtga accgtcagat cgcctggaga
      721 cgccatccac gctgttttga cctccataga agacaccggg accgatccag cctccggact
      781 ctagcgttta aacttaagct tcgccaccat gggatccccg gaagatgaga tccagcaact
      841 ggaagaagaa atcgctcagc tggaacagaa aaacgcagcg ctgaaagaga aaaaccaggc
      901 gctgaaatac ggtagtagtg gctcgagtaa atcagtctcc tccctcgtgt ccgaatcctc
      961 tcacatcgtt cacaaggaag ataccagttt ctggcagcat tggattacca caaaagacgg
     1021 gcagtgcggc tcaccgctgg tgtctatcat tgacgggaac attctgggaa tccactccct
     1081 gacacacacc actaacggct caaactattt cgttgagttc ccagagaagt tcgtagctac
     1141 atatctggac gctgcggacg gctggtgcaa gaactggaaa ttcaacgcgg acaagatttc
     1201 atggggtagt ttcacgcttg tcgaagacgc gcccgaggac gactttatgg cgaaaaaaac
     1261 cgtggctgct attatggata gctagctcga ggagggcccg aacaaaaact catctcagaa
     1321 gaggatctga atagcgccgt cgaccatcat cataatcatc attgagttta aacccgctga
     1381 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     1441 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     1501 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     1561 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggagatc
     1621 tatcgatgtt ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     1681 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     1741 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     1801 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     1861 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     1921 gatggttcac gtacctagaa gttcctattc cgaagttcct attctctaga aagtatagga
     1981 acttccttgg ccaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
     2041 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
     2101 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
     2161 tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg
     2221 cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt
     2281 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag
     2341 gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga
     2401 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
     2461 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
     2521 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     2581 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     2641 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     2701 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     2761 ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     2821 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     2881 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg
     2941 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     3001 agggcaaagg aatagcacgt actacgagat ttcgattcca ccgccgcctt ctatgaaagg
     3061 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     3121 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     3181 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3241 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     3301 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3361 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3421 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3481 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3541 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3601 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3661 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3721 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3781 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3841 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3901 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3961 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4021 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4081 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4141 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4201 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4261 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4321 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4381 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4441 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4501 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4561 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4621 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4681 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4741 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4801 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4861 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4921 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4981 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5041 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5101 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5161 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5221 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5281 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5341 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5401 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5461 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     5521 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     5581 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     5641 tgaagaatc
//
