LOCUS       sequence-398270-        7087 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express 40 Angstrom 12TMD de novo protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7087)
  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 7087)
  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: pJPT028-40 Angstrom 12TMD de novo_mRFP1_3x FLAG
FEATURES             Location/Qualifiers
     source          1..7087
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             262..936
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             262..936
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(392..410)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     660..679
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     854..877
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             952..1017
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1070..1191
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1081..1100)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1135..1154
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1306..1352
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1382..1518
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1563..1634
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1860..2448)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1940..1959)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2948..4157
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3994..4013
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4161..4760
                     /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(4161..4180)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4657..4677
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4787..4835
                     /note="synthetic polyadenylation signal"
     CDS             join(5101..5169,5170..5961)
                     /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(5319..5338)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5974..6110
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6262..6333
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6338..6384)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6402..6450
                     /note="synthetic polyadenylation signal"
     enhancer        6531..6834
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6836..7038
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6989..7009
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gacacacccg ccagcggccg cgccaccatg accaagaaaa tcatcatggt gctgatcctg
       61 ctgctgatca ttgctatgct gctgctggtg ttcctgctga tcgccaccgt ggtttccctg
      121 tggtggtcct ggaccgacga tgatcctagc gccatttctg aagccctggt ggccatgagc
      181 ctgatgctga ttgctgccag cctgctgatt atcgccatca gcaagctgct gaagtccaag
      241 aacggcggat ccggcggctc catggcctcc tccgaggacg tcatcaagga gttcatgcgc
      301 ttcaaggtgc gcatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
      361 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
      421 ctgcccttcg cctgggacat cctgtcccct cagttccagt acggctccaa ggcctacgtg
      481 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
      541 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
      601 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
      661 cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcggat gtaccccgag
      721 gacggcgccc tgaagggcga gatcaagatg aggctgaagc tgaaggacgg cggccactac
      781 gacgccgagg tcaagaccac ctacatggcc aagaagcccg tgcagctgcc cggcgcctac
      841 aagaccgaca tcaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
      901 tacgagcgcg ccgagggccg ccactccacc ggcgcctccg gctccgaatt cgattacaag
      961 gaccacgatg gcgactataa ggatcacgac atcgactaca aggacgatga cgacaagtga
     1021 gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat aagatacatt
     1081 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     1141 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     1201 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     1261 taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca taaccccttg
     1321 gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt gctatctacc
     1381 ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg
     1441 ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta
     1501 ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa
     1561 gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt tcatcaaact
     1621 ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt aaccagtcaa
     1681 gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt
     1741 caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt catttaaatc
     1801 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1861 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1921 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1981 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2041 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2101 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2161 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2221 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2281 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2341 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2401 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2461 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2521 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2581 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2641 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa atttccgaac
     2701 tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct tgcaggctac
     2761 ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta ttgcttggca
     2821 gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc acccgagaga
     2881 agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc gaaatcgggg
     2941 cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cgtcacggcg
     3001 agcgctgcca cgtcagacga agggcgcagg agcgtcctga tccttccgcc cggacgctca
     3061 ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag cagaaggaca
     3121 ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag agcggaacag
     3181 gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac
     3241 gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg cagccgggat
     3301 ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt agcgggctgc
     3361 tgggctggcc ggggctttcg tggccgccgg gccgctcggt gggacggaag cgtgtggaga
     3421 gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg ggggttgggg
     3481 ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg
     3541 ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc aaggtcttga
     3601 gcccttcgct aatgcgggaa agctcttatt cgggtgagat gggctgggca ccatctgggg
     3661 accctgacgt gaagtttgtc actgactgga gaactcggtt tgtcgtctgt tgcgggggcg
     3721 gcagttatgg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc
     3781 gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac ctgccggtag
     3841 gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg taggctctcc
     3901 tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt cagtttcttt
     3961 ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga actatgcgct
     4021 cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg taatcatttg
     4081 ggtcaatatg taattttcag tgttagactt gtaaattgtc cgctaaattc tggccgtttt
     4141 tggctttttt gttagacaac atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     4201 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     4261 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     4321 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     4381 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     4441 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggag ggcctcctgg
     4501 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     4561 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcc gccgagcgcg
     4621 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     4681 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     4741 cccgcaagcc cggtgcctag gctagtatgt aagcctagtc ttagataata aaatcgctat
     4801 ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa cgctaggcgc gcctggtgta
     4861 ccgagaacga tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc
     4921 agtcggaatc cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg
     4981 tcacggcagc gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat
     5041 aatcgagtcc tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc
     5101 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5161 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg
     5221 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5281 gaagaacgct ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5341 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5401 gttgagtatt caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5461 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt
     5521 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5581 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5641 cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct
     5701 tcccggcaac agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5761 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5821 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5881 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5941 tcactgatta agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat
     6001 gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc
     6061 caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa gatcccgaat
     6121 cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc
     6181 gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt
     6241 ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc
     6301 tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt
     6361 tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca
     6421 catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc
     6481 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     6541 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     6601 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     6661 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     6721 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     6781 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catgctgatg
     6841 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     6901 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     6961 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     7021 gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca
     7081 tccactc
//
