LOCUS       sequence-361444-        8586 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For exrpression of mCardinal targeted to peroxisomes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8586)
  AUTHORS   Haimovich G, Dasgupta S, Ravi AG, Gerst JE
  TITLE     Complementation of a human disease phenotype in vitro by 
            intercellular mRNA transfer
  JOURNAL   bioRxiv 2024.11.06.622258
REFERENCE   2  (bases 1 to 8586)
  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: "bioRxiv 
            2024.11.06.622258"
COMMENT     Sequence Label: pHAGE-mCardinal-SKL
FEATURES             Location/Qualifiers
     source          1..8586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(67..135,136..927)
                     /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(285..304)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1098..1686
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1587..1606
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1840..1857
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1974..1995
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2010..2015,2016..2033,2034..2040)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2048..2064
                     /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)."
     primer_bind     2053..2075
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2072..2088
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2072..2088
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2107..2127
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2109..2127
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        2228..2607
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2608..2811
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2762..2782
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2826..3006
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3053..3178
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3671..3904
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4089..4133
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    4400..4517
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4574..5785
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5622..5641
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      5819..5828
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5825..6556
                     /codon_start=1
                     /product="far-red fluorescent protein derived from 
                     mNeptune, with further red-shifted spectra"
                     /label=mCardinal
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTTEGEGKPYEGTQTQ
                     RIKVVEGGPLPFAFDILATCFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTVTQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEATTETLYPADGGLEGRCD
                     MALKLVGGGHLHCNLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADNETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     misc_feature    6584..7172
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6637..6657)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7055..7066)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7259..7492
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7564..7685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7601..7620)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7655..7674
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7725..7860
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7880..7899)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7881..7899)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7909..7926)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7909..7925)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7918..7940)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8067..8522
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8154..8173)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8364..8385
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        join(8548..8586,1..66)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
       61 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
      121 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
      181 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
      241 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
      301 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
      361 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
      421 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
      481 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
      541 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
      601 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
      661 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
      721 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
      781 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
      841 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
      901 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
      961 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1021 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1081 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1141 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1201 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1261 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1321 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1381 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1441 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1501 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1561 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1621 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     1681 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     1741 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     1801 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     1861 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     1921 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     1981 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     2041 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     2101 aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc ttggccattg
     2161 catacgttgt atccatatca taatatgtac atttatattg gctcatgtcc aacattaccg
     2221 ccatgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     2281 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     2341 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     2401 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     2461 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     2521 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     2581 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     2641 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     2701 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     2761 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
     2821 aaccggggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     2881 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     2941 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     3001 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
     3061 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
     3121 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
     3181 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
     3241 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
     3301 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
     3361 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
     3421 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
     3481 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag
     3541 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
     3601 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
     3661 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
     3721 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
     3781 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     3841 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     3901 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     3961 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     4021 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     4081 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     4141 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     4201 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     4261 attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag
     4321 gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat
     4381 ctcgacggta tcggttaact tttaaaagaa aaggggggat tggggggtac agtgcagggg
     4441 aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta
     4501 caaaaattca aaattttatc gatcacgaga ctagcctcga gaagcttgat taacccgtgt
     4561 cggctccaga tctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcctc
     4621 acggcgagcg ctgccacgtc agacgaaggg cgcagcgagc gtcctgatcc ttccgcccgg
     4681 acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca gtatcagcag
     4741 aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt tccagagagc
     4801 ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc tccgtggggc
     4861 ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag
     4921 ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc
     4981 gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt
     5041 gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg
     5101 gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg
     5161 aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag
     5221 gtcttgaggc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctggggcacc
     5281 atctggggac cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg
     5341 cgggggcggc agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg
     5401 ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacct
     5461 gccggtaggt gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta
     5521 ggctctcctg aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca
     5581 gtttctttgg tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac
     5641 tatgcgctcg gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta
     5701 atcatttggg tcaatatgta attttcagtg ttagactagt aaattgtccg ctaaattctg
     5761 gccgtttttg gcttttttgt tagacgaagc ttgggctgca ggtcgagatc caccggtcgc
     5821 caccatggtg agcaagggcg aggagctgat caaggagaac atgcacatga agctgtacat
     5881 ggaaggcacc gtgaacaacc accacttcaa gtgcaccacc gaaggggagg gcaagcccta
     5941 cgagggcacc cagacccaga ggattaaggt ggtggaggga ggccccctgc cgttcgcatt
     6001 cgacatcctg gccacctgct ttatgtacgg gagcaagacc ttcatcaacc acacccaggg
     6061 catccccgat ttctttaagc agtccttccc tgagggcttc acatgggaga gagtcaccac
     6121 atacgaagac gggggcgtgc ttaccgttac ccaggacacc agcctccagg acggctgctt
     6181 gatctacaac gtcaagctca gaggggtgaa cttcccatcc aacggccctg tgatgcagaa
     6241 gaaaacactc ggctgggagg ccaccaccga gaccctgtac cccgctgacg gcggcctgga
     6301 aggcagatgc gacatggccc tgaagctcgt gggcgggggc cacctgcact gcaacctgaa
     6361 gaccacatac agatccaaga aacccgctaa gaacctcaag atgcccggcg tctactttgt
     6421 ggaccgcaga ctggaaagaa tcaaggaggc cgacaatgag acctacgtcg agcagcacga
     6481 ggtggctgtg gccagatact gcgacctccc tagcaaactg gggcacaaac ttaatggcat
     6541 ggacgagctg tacaagagca agctctaaag cggccgcgtc gacaatcaac ctctggatta
     6601 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     6661 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     6721 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     6781 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     6841 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     6901 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     6961 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     7021 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     7081 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     7141 gagtcggatc tccctttggg ccgcctcccc gcctggaatt cgagctcggt acctttaaga
     7201 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     7261 gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc
     7321 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     7381 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     7441 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt
     7501 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
     7561 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     7621 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     7681 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc
     7741 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     7801 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     7861 tagggacgta cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt
     7921 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     7981 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     8041 gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     8101 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     8161 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     8221 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     8281 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     8341 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     8401 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     8461 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     8521 ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     8581 cattca
//
