LOCUS       sequence-399273-        5125 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DsRed-Express2 reporter regulated by ZF6 synTF promoter (12x compact
            binding sites) in TUPV1 for mMoClo golden gate-based assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5125)
  AUTHORS   Peruzzi JA, Gunnels TF, Edelstein HI, Lu P, Baker D, Leonard JN, 
            Kamat NP
  TITLE     Enhancing extracellular vesicle cargo loading and functional 
            delivery by engineering protein-lipid interactions.
  JOURNAL   Nat Commun. 2024 Jul 4;15(1):5618. doi: 10.1038/s41467-024-49678-z.
  PUBMED    38965227
REFERENCE   2  (bases 1 to 5125)
  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: "2024-07-4"; volume: "15(1):5618. doi"; pages: " 
            10.1038/s41467-024-49678-z"
COMMENT     Sequence Label: pHIE324-ZF6x12-C DsRed-Express2 in TUPV1
FEATURES             Location/Qualifiers
     source          1..5125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      304..313
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             310..987
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(490..510)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     902..925
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1032..1049)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1038..1262
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2031..2459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2118..2137)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2328..2349
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2482..2678
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2529..2664
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2591..2610
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2798..2919
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2835..2854)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2889..2908
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2968..2984)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2968..2984)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2981..3003)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2992..3008
                     /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(3016..3022,3023..3040,3041..3046))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3061..3082
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3199..3216)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3370..3958)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3450..3469)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4129..4920,4921..4989))
                     /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     4752..4771
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4990..5094)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgaag acggtgccaa ggtcttgaca aacgagtgct agtacaagat
       61 ctgcgagggt gtaggggtta atggtgtagg ggtaattggt gtaggggtct agggtgtagg
      121 ggtaatgggt gtaggggttt aaggtgtagg ggtagcaggt gtaggggttc gtggtgtagg
      181 ggttataggt gtaggggtaa ttggtgtagg ggtatgtggt gtaggggtaa cgggtgtagg
      241 ggtacgctct ctagagggta tataatgggg gccactagtc tactaccaga gctcatcgct
      301 agcgccacca tggatagcac tgagaacgtc atcaagccct tcatgcgctt caaggtgcac
      361 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggcaagccc
      421 tacgagggca cccagaccgc caagctccag gtgaccaagg gcggccccct gcccttcgcc
      481 tgggacatcc tgtcccccca gttccagtac ggctccaagg tgtacgtgaa gcaccccgcc
      541 gacatccccg actacaagaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
      601 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctcca ggacggcacc
      661 ttcatctacc acgtgaagtt catcggcgtg aacttcccct ccgacggccc cgtaatgcag
      721 aagaaaactc tgggctggga gccctccacc gagcgcctgt acccccgcga cggcgtgctg
      781 aagggcgaga tccacaaggc gctgaagctg aagggcggcg gccactacct ggtggagttc
      841 aagtcaatct acatggccaa gaagcccgtg aagctgcccg gctactacta cgtggactcc
      901 aagctggaca tcacctccca caacgaggac tacaccgtgg tggagcagta cgagcgcgcc
      961 gaggcccgcc accacctgtt ccagtaggcg gccgctcgag tctagagggc ccgtttaaac
     1021 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     1081 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     1141 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     1201 cagcaagggg gaggattggg aagtcaatag caggcatgct ggggatgcgg tgggctctat
     1261 ggcttctgag gcaggtcgag ggtacatgcg ccttactcct tgtgagcgca tcgcgataag
     1321 ctctagaggg acagcccccc cccaaagccc ccagggatgt aattacgtcc ctcccccgct
     1381 agggggcagc agcgagccgc ccggggctcc gctccggtcc ggcgctcccc ccgcatcccc
     1441 gagccggcag cgtgcgggga cagcccgggc acggggaagg tggcacggga tcgctttcct
     1501 ctgaacgctt ctcgctgctc tttgagcctg cagacacctg gggggatacg gggaaaaagc
     1561 tttaggctga aagagagatt tagaatgaca gaatcataga acggcctgcg tacggatggg
     1621 gctctagctc tagagcttga tatcgaattc ctgcagcccc gctagaggga cagccccccc
     1681 ccaaagcccc cagggatgta attacgtccc tcccccgcta gggggcagca gcgagccgcc
     1741 cggggctccg ctccggtccg gcgctccccc cgcatccccg agccggcagc gtgcggggac
     1801 agcccgggca cggggaaggt ggcacgggat cgctttcctc tgaacgcttc tcgctgctct
     1861 ttgagcctgc agacacctgg ggggatacgg ggaaaaagct ttaggctgaa agagagattt
     1921 agaatgacag aatcatagaa cggcctgggc gcgcctggtg gcgaacgatc aacagacggg
     1981 agagggcaaa agtcttcgct gacagcgacc ggactctagg gggtatcccc acgcgccctg
     2041 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2101 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2161 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2221 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2281 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2341 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2401 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     2461 attctgtgga atgtgtgtca gttaggctca attagtcagc aaccatagtc ccgcccctaa
     2521 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2581 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     2641 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     2701 ccattttcgg aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     2761 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     2821 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2881 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     2941 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     3001 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3061 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3121 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3181 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3241 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3301 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3361 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3421 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3481 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3541 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3601 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3661 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3721 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3781 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3841 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3901 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3961 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4021 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4081 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4141 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4201 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4261 gataccgcga gatccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4321 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4381 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4441 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4501 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4561 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4621 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4681 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4741 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4801 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4861 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4921 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4981 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5041 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5101 tccccgaaaa gtgccacctg acgtc
//
