LOCUS       sequence-355669-        5751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ER-associated uncaged-nTVMVP with AP4 leucine zipper.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5751)
  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 5751)
  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-AP4
FEATURES             Location/Qualifiers
     source          1..5751
                     /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             3097..3126
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3182..3199)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3188..3412
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3584..3603)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3735..3782
                     /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             3790..4812
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3790..4812
                     /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    4942..5063
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4979..4998)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5033..5052
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5112..5128)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5112..5128)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5125..5147)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5136..5152
                     /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(5160..5166,5167..5184,5185..5190))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5205..5226
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5343..5360)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5514..5751,1..351))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5594..5613)
                     /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 aggcagttca cccgaggacg agcttgctgc
     3001 taatgaagag gagttgcagc aaaatgaaca aaagttggct caaattaagc aaaaacttca
     3061 agctatcaaa tacggtagct agctcgagga gggcccgaac aaaaactcat ctcagaagag
     3121 gatctgaata gcgccgtcga ccatcatcat aatcatcatt gagtttaaac ccgctgatca
     3181 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3241 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3301 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3361 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggagatctat
     3421 cgatgttctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3481 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3541 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3601 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3661 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3721 ggttcacgta cctagaagtt cctattccga agttcctatt ctctagaaag tataggaact
     3781 tccttggcca aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag
     3841 ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc
     3901 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac
     3961 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt
     4021 gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc
     4081 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc
     4141 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg
     4201 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat
     4261 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc
     4321 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat
     4381 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc
     4441 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg
     4501 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga
     4561 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg
     4621 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga
     4681 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc
     4741 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg
     4801 gcaaaggaat agcacgtact acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4861 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4921 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4981 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5041 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5101 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5161 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5221 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5281 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5341 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5401 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5461 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5521 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5581 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5641 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5701 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg c
//
