LOCUS       sequence-361442-        8586 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For exrpression of mNeptune2 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-mNeptune2-SKL
FEATURES             Location/Qualifiers
     source          1..8586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..107
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(108..176,177..968)
                     /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(326..345)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1139..1727
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1628..1647
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1881..1898
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2015..2036
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2051..2056,2057..2074,2075..2081)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2089..2105
                     /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     2094..2116
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2113..2129
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2113..2129
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2148..2168
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2150..2168
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        2269..2648
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2649..2852
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2803..2823
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2867..3047
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3094..3219
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3712..3945
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4130..4174
                     /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    4441..4558
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4615..5826
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5663..5682
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      5860..5869
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5866..6597
                     /codon_start=1
                     /product="far-red fluorescent protein with spectra similar 
                     to those of mNeptune but improved brightness"
                     /label=mNeptune2
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTG
                     RIKVVEGGPLPFAFDILATCFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTVTQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRCD
                     MALKLVGGGHLHCNLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADNETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     misc_feature    6625..7213
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6678..6698)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7096..7107)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7300..7533
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7605..7726
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7642..7661)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7696..7715
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7766..7901
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7921..7940)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7922..7940)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7950..7967)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7950..7966)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7959..7981)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8108..8563
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8195..8214)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8405..8426
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
       61 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
      121 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
      181 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
      241 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
      301 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
      361 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
      421 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
      481 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
      541 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
      601 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
      661 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
      721 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
      781 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
      841 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
      901 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
      961 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     1021 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     1081 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1141 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1201 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1261 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     1321 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     1381 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     1441 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     1501 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     1561 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     1621 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     1681 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     1741 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     1801 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     1861 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     1921 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     1981 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     2041 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     2101 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
     2161 actaaaggga acaaaagctg gagctgcaag cttggccatt gcatacgttg tatccatatc
     2221 ataatatgta catttatatt ggctcatgtc caacattacc gccatgttga cattgattat
     2281 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     2341 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     2401 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     2461 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     2521 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     2581 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     2641 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     2701 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     2761 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     2821 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccggggt ctctctggtt
     2881 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     2941 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     3001 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac
     3061 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct
     3121 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact
     3181 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt
     3241 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa
     3301 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa
     3361 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca
     3421 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata
     3481 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag
     3541 accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa
     3601 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
     3661 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
     3721 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
     3781 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
     3841 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
     3901 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
     3961 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
     4021 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
     4081 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
     4141 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
     4201 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
     4261 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
     4321 gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg
     4381 agagagagac agagacagat ccattcgatt agtgaacgga tctcgacggt atcggttaac
     4441 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
     4501 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttat
     4561 cgatcacgag actagcctcg agaagcttga ttaacccgtg tcggctccag atctggcctc
     4621 cgcgccgggt tttggcgcct cccgcgggcg cccccctcct cacggcgagc gctgccacgt
     4681 cagacgaagg gcgcagcgag cgtcctgatc cttccgcccg gacgctcagg acagcggccc
     4741 gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg
     4801 acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt
     4861 agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat
     4921 ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt
     4981 tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg ggctggccgg
     5041 ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg
     5101 ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg ggttgggggg agcgcagcaa
     5161 aatggcggct gttcccgagt cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt
     5221 tgaaacaagg tggggggcat ggtgggcggc aagaacccaa ggtcttgagg ccttcgctaa
     5281 tgcgggaaag ctcttattcg ggtgagatgg gctggggcac catctgggga ccctgacgtg
     5341 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     5401 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     5461 tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg tgtgcggtag
     5521 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     5581 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     5641 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     5701 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     5761 aattttcagt gttagactag taaattgtcc gctaaattct ggccgttttt ggcttttttg
     5821 ttagacgaag cttgggctgc aggtcgagat ccaccggtcg ccaccatggt gtctaagggc
     5881 gaagagctga ttaaggagaa catgcacatg aagctgtaca tggagggcac cgtgaacaac
     5941 caccacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac ccagaccggc
     6001 agaatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct ggctacctgc
     6061 ttcatgtacg gcagcaagac cttcatcaac cacacccagg gcatccccga tttctttaag
     6121 cagtccttcc ctgagggctt cacatgggag agagtcacca catacgaaga cgggggcgtg
     6181 cttaccgtta cccaggacac cagcctccag gacggctgct tgatctacaa cgtcaagctc
     6241 agaggggtga acttcccatc caacggccct gtgatgcaga agaaaacact cggctgggag
     6301 gccagtaccg agaccctgta ccccgctgac ggcggcctgg aaggcagatg cgacatggcc
     6361 ctgaagctcg tgggcggggg ccacctgcac tgcaacctga agaccacata cagatccaag
     6421 aaacccgcta agaacctcaa gatgcccggc gtctactttg tggaccgcag actggaaaga
     6481 atcaaggagg ccgacaatga gacctacgtc gagcagcacg aggtggctgt ggccagatac
     6541 tgcgacctcc ctagcaaact ggggcacaaa cttaatggca tggacgagct gtacaagagc
     6601 aagctctaaa gcggccgcgt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     6661 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     6721 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     6781 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     6841 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     6901 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     6961 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     7021 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     7081 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     7141 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     7201 gccgcctccc cgcctggaat tcgagctcgg tacctttaag accaatgact tacaaggcag
     7261 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     7321 aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     7381 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     7441 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     7501 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     7561 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     7621 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     7681 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     7741 agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     7801 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     7861 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg
     7921 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     7981 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     8041 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     8101 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     8161 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     8221 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     8281 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     8341 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     8401 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     8461 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     8521 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     8581 gaaatg
//
