LOCUS       sequence-398262-        7354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express ""G"" synTF (ZF6).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7354)
  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 7354)
  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: pJPT010-ZF6 1x FLAG-synTF-G
FEATURES             Location/Qualifiers
     source          1..7354
                     /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             31..54
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             70..90
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             523..879
                     /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"
     polyA_signal    1337..1458
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1348..1367)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1402..1421
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1573..1619
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1649..1785
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1830..1901
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2127..2715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2207..2226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3215..4424
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4261..4280
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4428..5027
                     /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(4428..4447)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4924..4944
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5054..5102
                     /note="synthetic polyadenylation signal"
     CDS             join(5368..5436,5437..6228)
                     /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(5586..5605)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6241..6377
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6529..6600
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6605..6651)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6669..6717
                     /note="synthetic polyadenylation signal"
     enhancer        6798..7101
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7103..7305
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7256..7276
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gacacacccg ccagcggccg cgccaccatg gactacaagg acgatgacga caaggaattc
       61 ggcggctccc ctaagaaaaa gcgcaaagtc tccggaagcc agtacctgcc tgacaccgac
      121 gaccggcaca gaatcgagga aaagcggaag cggacctacg agacattcaa gagcatcatg
      181 aagaagtccc cattcagcgg ccccaccgat cctagacctc cacctagaag aatcgccgtg
      241 cctagcagat ccagcgcctc tgtgcctaaa cctgctcctc agccttatcc tttcaccagc
      301 agcctgagca ccatcaacta cgacgagttc cccacaatgg tgttccccag cggacagatc
      361 agccaggctt ctgctcttgc tccagctcct cctcaggttc tgcctcaagc tcctgctccg
      421 gctccagcac cagctatggt ttctgctttg gcccaggctc ctgcacctgt gcctgttctt
      481 gctcctggac cacctcaggc tgttgctcca ccagctccta aacctacaca ggccggcgag
      541 ggaacactgt ctgaagccct gctgcaactc cagttcgacg acgaggatct gggagcactg
      601 ctgggcaata gcacagaccc tgccgtgttt accgatctgg ccagcgtgga caacagcgag
      661 tttcagcagc tcctgaacca gggcatccct gtggctcctc acaccacaga gcccatgctg
      721 atggaatacc ccgaggccat caccagactg gtcaccggcg ctcaaagacc tccagatcct
      781 gcaccagcac ctcttggagc acctggcctg cctaatggac tgctgagcgg agatgaggac
      841 ttcagctcta tcgccgacat ggatttcagc gccctgctcg gttccggtgg tggcgggtct
      901 ggaggaggag gtagtggtgg aggtggctct ggtaccgcta gacccggcga aagacctttc
      961 cagtgccgga tctgcatgag gaacttcagc aagggcgaga gactcgtgcg gcacaccaga
     1021 acacacacag gcgagaagcc cttccagtgt agaatctgta tgcgcaactt cagccggatg
     1081 gacaacctga gcacccacct gagaacccat accggggaga agccatttca atgccgcatc
     1141 tgtatgagaa atttttcccg gaaggacgcc ctgaaccggc acctgaaaac acacctgaga
     1201 ggcagcggct ccggatccgg tggtagcttt cgtagtgatg gcaaaaagaa aaagaagaaa
     1261 tccaagacca aatgccagct gctgtgagct agcttgactg actgagatac agcgtacctt
     1321 cagctcacag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     1381 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     1441 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     1501 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct
     1561 ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt
     1621 gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac
     1681 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     1741 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc
     1801 tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac
     1861 taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg
     1921 gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt
     1981 ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt
     2041 tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac
     2101 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2161 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2221 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2281 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2341 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2401 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2461 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2521 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2581 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2641 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2701 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2761 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2821 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2881 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2941 tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc
     3001 tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg
     3061 gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa
     3121 tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc
     3181 gagatccgag gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt tttggcgcct
     3241 cccgcgggcg cccccctcgt cacggcgagc gctgccacgt cagacgaagg gcgcaggagc
     3301 gtcctgatcc ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct
     3361 tagaacccca gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac
     3421 tggttttctt tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg
     3481 cggagggatc tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg
     3541 cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg
     3601 atcgtcactt ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc
     3661 gctcggtggg acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc
     3721 aaggttgccc tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc
     3781 ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg
     3841 gtgggcggca agaacccaag gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg
     3901 gtgagatggg ctgggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa
     3961 ctcggtttgt cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc
     4021 gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata
     4081 atgcagggtg gggccacctg ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc
     4141 agggttcggg cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg
     4201 agggataagt gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc
     4261 tgaagctccg gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt
     4321 aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagacttgta
     4381 aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacaacatg accgagtaca
     4441 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     4501 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     4561 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     4621 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     4681 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     4741 cgcagcaaca gatggagggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     4801 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     4861 tccccggagt ggaggccgcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     4921 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     4981 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctaggct agtatgtaag
     5041 cctagtctta gataataaaa tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg
     5101 tgtgtaacgc taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg
     5161 atccatatcg ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca
     5221 atcgtcagat attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga
     5281 tattgatagt ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa
     5341 gagacaggat cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt
     5401 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5461 aaagatgctg aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc
     5521 ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa
     5581 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5641 cgcatacact attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt
     5701 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5761 gcggccaact tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac
     5821 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5881 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta
     5941 ttaactggcg aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg
     6001 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6061 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6121 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6181 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt
     6241 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     6301 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     6361 cagaaattta tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc
     6421 cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct
     6481 cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca
     6541 ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc
     6601 aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt
     6661 gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc
     6721 taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc
     6781 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     6841 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     6901 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     6961 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     7021 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     7081 ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag
     7141 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     7201 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     7261 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
     7321 cagatcagat ctttgtcgat cctaccatcc actc
//
