LOCUS       sequence-398260-        7387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express ""PM"" synTF (ZF6).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7387)
  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 7387)
  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: pJPT009-ZF6 PM-synTF-1x FLAG
FEATURES             Location/Qualifiers
     source          1..7387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      22..31
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             145..165
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             598..954
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             1294..1317
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1370..1491
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1381..1400)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1435..1454
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1606..1652
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1682..1818
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1863..1934
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2160..2748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2240..2259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3248..4457
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4294..4313
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4461..5060
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4461..4480)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4957..4977
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5087..5135
                     /note="synthetic polyadenylation signal"
     CDS             join(5401..5469,5470..6261)
                     /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     complement(5619..5638)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6274..6410
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6562..6633
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6638..6684)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6702..6750
                     /note="synthetic polyadenylation signal"
     enhancer        6831..7134
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7136..7338
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7289..7309
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gacacacccg ccagcggccg cgccaccatg ggctgcatca agagcaagcg gaaggacaag
       61 gacctggaac tgaagctgag aatcctgcag agcaccgtgc ctagagctag agatcctcca
      121 gtggccacag aattcggcgg ctcccctaag aaaaagcgca aagtctccgg aagccagtac
      181 ctgcctgaca ccgacgaccg gcacagaatc gaggaaaagc ggaagcggac ctacgagaca
      241 ttcaagagca tcatgaagaa gtccccattc agcggcccca ccgatcctag acctccacct
      301 agaagaatcg ccgtgcctag cagatccagc gcctctgtgc ctaaacctgc tcctcagcct
      361 tatcctttca ccagcagcct gagcaccatc aactacgacg agttccccac aatggtgttc
      421 cccagcggac agatcagcca ggcttctgct cttgctccag ctcctcctca ggttctgcct
      481 caagctcctg ctccggctcc agcaccagct atggtttctg ctttggccca ggctcctgca
      541 cctgtgcctg ttcttgctcc tggaccacct caggctgttg ctccaccagc tcctaaacct
      601 acacaggccg gcgagggaac actgtctgaa gccctgctgc aactccagtt cgacgacgag
      661 gatctgggag cactgctggg caatagcaca gaccctgccg tgtttaccga tctggccagc
      721 gtggacaaca gcgagtttca gcagctcctg aaccagggca tccctgtggc tcctcacacc
      781 acagagccca tgctgatgga ataccccgag gccatcacca gactggtcac cggcgctcaa
      841 agacctccag atcctgcacc agcacctctt ggagcacctg gcctgcctaa tggactgctg
      901 agcggagatg aggacttcag ctctatcgcc gacatggatt tcagcgccct gctcggttcc
      961 ggtggtggcg ggtctggagg aggaggtagt ggtggaggtg gctctggtac cgctagaccc
     1021 ggcgaaagac ctttccagtg ccggatctgc atgaggaact tcagcaaggg cgagagactc
     1081 gtgcggcaca ccagaacaca cacaggcgag aagcccttcc agtgtagaat ctgtatgcgc
     1141 aacttcagcc ggatggacaa cctgagcacc cacctgagaa cccataccgg ggagaagcca
     1201 tttcaatgcc gcatctgtat gagaaatttt tcccggaagg acgccctgaa ccggcacctg
     1261 aaaacacacc tgagaggcag cggctccgga tccgactaca aggacgatga cgacaagtga
     1321 gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat aagatacatt
     1381 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     1441 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     1501 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     1561 taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca taaccccttg
     1621 gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt gctatctacc
     1681 ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg
     1741 ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta
     1801 ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa
     1861 gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt tcatcaaact
     1921 ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt aaccagtcaa
     1981 gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt
     2041 caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt catttaaatc
     2101 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2161 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2221 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2281 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2341 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2401 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2461 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2521 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2581 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2641 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2701 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2761 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2821 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2881 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2941 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa atttccgaac
     3001 tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct tgcaggctac
     3061 ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta ttgcttggca
     3121 gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc acccgagaga
     3181 agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc gaaatcgggg
     3241 cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cgtcacggcg
     3301 agcgctgcca cgtcagacga agggcgcagg agcgtcctga tccttccgcc cggacgctca
     3361 ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag cagaaggaca
     3421 ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag agcggaacag
     3481 gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac
     3541 gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg cagccgggat
     3601 ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt agcgggctgc
     3661 tgggctggcc ggggctttcg tggccgccgg gccgctcggt gggacggaag cgtgtggaga
     3721 gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg ggggttgggg
     3781 ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg
     3841 ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc aaggtcttga
     3901 gcccttcgct aatgcgggaa agctcttatt cgggtgagat gggctgggca ccatctgggg
     3961 accctgacgt gaagtttgtc actgactgga gaactcggtt tgtcgtctgt tgcgggggcg
     4021 gcagttatgg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc
     4081 gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac ctgccggtag
     4141 gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg taggctctcc
     4201 tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt cagtttcttt
     4261 ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga actatgcgct
     4321 cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg taatcatttg
     4381 ggtcaatatg taattttcag tgttagactt gtaaattgtc cgctaaattc tggccgtttt
     4441 tggctttttt gttagacaac atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     4501 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     4561 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     4621 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     4681 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     4741 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggag ggcctcctgg
     4801 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     4861 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcc gccgagcgcg
     4921 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     4981 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     5041 cccgcaagcc cggtgcctag gctagtatgt aagcctagtc ttagataata aaatcgctat
     5101 ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa cgctaggcgc gcctggtgta
     5161 ccgagaacga tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc
     5221 agtcggaatc cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg
     5281 tcacggcagc gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat
     5341 aatcgagtcc tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc
     5401 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5461 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg
     5521 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5581 gaagaacgct ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5641 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5701 gttgagtatt caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5761 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt
     5821 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5881 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5941 cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct
     6001 tcccggcaac agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6061 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     6121 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6181 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6241 tcactgatta agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat
     6301 gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc
     6361 caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa gatcccgaat
     6421 cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc
     6481 gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt
     6541 ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc
     6601 tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt
     6661 tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca
     6721 catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc
     6781 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     6841 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     6901 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     6961 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     7021 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     7081 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catgctgatg
     7141 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     7201 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     7261 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     7321 gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca
     7381 tccactc
//
