LOCUS       sequence-398264-        7288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express ""soluble"" synTF (ZF6).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7288)
  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 7288)
  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: pJPT007-ZF6 synTF-1x FLAG
FEATURES             Location/Qualifiers
     source          1..7288
                     /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             46..66
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             499..855
                     /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             1195..1218
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1271..1392
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1282..1301)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1336..1355
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1507..1553
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1583..1719
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1764..1835
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2061..2649)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2141..2160)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3149..4358
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4195..4214
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4362..4961
                     /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(4362..4381)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4858..4878
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4988..5036
                     /note="synthetic polyadenylation signal"
     CDS             join(5302..5370,5371..6162)
                     /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(5520..5539)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6175..6311
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6463..6534
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6539..6585)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6603..6651
                     /note="synthetic polyadenylation signal"
     enhancer        6732..7035
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7037..7239
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7190..7210
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gacacacccg ccagcggccg cgccaccatg gaattcggcg gctcccctaa gaaaaagcgc
       61 aaagtctccg gaagccagta cctgcctgac accgacgacc ggcacagaat cgaggaaaag
      121 cggaagcgga cctacgagac attcaagagc atcatgaaga agtccccatt cagcggcccc
      181 accgatccta gacctccacc tagaagaatc gccgtgccta gcagatccag cgcctctgtg
      241 cctaaacctg ctcctcagcc ttatcctttc accagcagcc tgagcaccat caactacgac
      301 gagttcccca caatggtgtt ccccagcgga cagatcagcc aggcttctgc tcttgctcca
      361 gctcctcctc aggttctgcc tcaagctcct gctccggctc cagcaccagc tatggtttct
      421 gctttggccc aggctcctgc acctgtgcct gttcttgctc ctggaccacc tcaggctgtt
      481 gctccaccag ctcctaaacc tacacaggcc ggcgagggaa cactgtctga agccctgctg
      541 caactccagt tcgacgacga ggatctggga gcactgctgg gcaatagcac agaccctgcc
      601 gtgtttaccg atctggccag cgtggacaac agcgagtttc agcagctcct gaaccagggc
      661 atccctgtgg ctcctcacac cacagagccc atgctgatgg aataccccga ggccatcacc
      721 agactggtca ccggcgctca aagacctcca gatcctgcac cagcacctct tggagcacct
      781 ggcctgccta atggactgct gagcggagat gaggacttca gctctatcgc cgacatggat
      841 ttcagcgccc tgctcggttc cggtggtggc gggtctggag gaggaggtag tggtggaggt
      901 ggctctggta ccgctagacc cggcgaaaga cctttccagt gccggatctg catgaggaac
      961 ttcagcaagg gcgagagact cgtgcggcac accagaacac acacaggcga gaagcccttc
     1021 cagtgtagaa tctgtatgcg caacttcagc cggatggaca acctgagcac ccacctgaga
     1081 acccataccg gggagaagcc atttcaatgc cgcatctgta tgagaaattt ttcccggaag
     1141 gacgccctga accggcacct gaaaacacac ctgagaggca gcggctccgg atccgactac
     1201 aaggacgatg acgacaagtg agctagcttg actgactgag atacagcgta ccttcagctc
     1261 acagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     1321 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     1381 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     1441 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg
     1501 gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct
     1561 cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     1621 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     1681 cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg
     1741 cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg
     1801 tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc
     1861 gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac
     1921 tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg
     1981 attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2041 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2101 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2161 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2221 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2281 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2341 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2401 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2461 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2521 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2581 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2641 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     2701 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     2761 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     2821 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2881 gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt
     2941 ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg
     3001 cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg
     3061 agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc
     3121 cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg
     3181 ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg
     3241 atccttccgc ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac
     3301 cccagtatca gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt
     3361 tctttccaga gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg
     3421 gatctccgtg gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc
     3481 tagttccgtc gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc
     3541 acttggtgag tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg
     3601 tgggacggaa gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt
     3661 gccctgaact gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat
     3721 ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc
     3781 ggcaagaacc caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga
     3841 tgggctgggc accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt
     3901 ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct
     3961 ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag
     4021 ggtggggcca cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt
     4081 cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat
     4141 aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc
     4201 tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac
     4261 cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt
     4321 ccgctaaatt ctggccgttt ttggcttttt tgttagacaa catgaccgag tacaagccca
     4381 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     4441 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     4501 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     4561 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     4621 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     4681 aacagatgga gggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     4741 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     4801 gagtggaggc cgccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     4861 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     4921 gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta ggctagtatg taagcctagt
     4981 cttagataat aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta
     5041 acgctaggcg cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat
     5101 atcgttgctt ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc
     5161 agatattgta ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga
     5221 tagtctgatc ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca
     5281 ggatcagcag gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt
     5341 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5401 gctgaagatc agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5461 atccttgaga gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg
     5521 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5581 cactattctc agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat
     5641 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5701 aacttacttc tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5761 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5821 gacgagcgtg acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact
     5881 ggcgaactac ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa
     5941 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6001 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6061 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6121 cagatcgctg agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg
     6181 gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt
     6241 cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa
     6301 tttatcctta agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt
     6361 ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag
     6421 ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct
     6481 ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca
     6541 aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta
     6601 cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg
     6661 ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca tagcccatat
     6721 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     6781 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     6841 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     6901 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     6961 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     7021 atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     7081 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     7141 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     7201 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
     7261 agatctttgt cgatcctacc atccactc
//
