LOCUS       sequence-456142-        5529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express N-terminal mCherry fused to human PHOSPHO1 in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5529)
  TITLE     PHOSPHO2 is PA phosphatase and PHOSPHO1 is PLC
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5529)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pmCherry-C1-PHOSPHO1
FEATURES             Location/Qualifiers
     source          1..5529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..57
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    181..302
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(218..237)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     272..291
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(309..764)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(446..467)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     658..677
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        791..895
                     /gene="bla"
                     /label=AmpR promoter
     promoter        897..1254
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        897..1088
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(920..940)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1105..1240
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1289..2083
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1289..2083
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1343..1362)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1953..1972
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2271..2290)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2315..2362
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2691..3279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3180..3199
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3454..3757
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3758..3961
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3912..3932
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4000..4009
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4006..4713
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4151..4169)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4419..4438
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 agatctcgag ctcaagcttc gaattctgca gtcgacggta ccgcgggccc gggatccacc
       61 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
      121 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
      181 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
      241 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
      301 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
      361 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
      421 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
      481 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
      541 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
      601 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
      661 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
      721 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
      781 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
      841 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
      901 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
      961 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     1021 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     1081 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     1141 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     1201 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     1261 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     1321 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     1381 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     1441 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     1501 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     1561 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     1621 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     1681 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     1741 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     1801 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     1861 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     1921 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     1981 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     2041 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     2101 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     2161 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     2221 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     2281 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     2341 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     2401 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     2461 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     2521 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     2581 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2641 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2701 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2761 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2821 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2881 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2941 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3001 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3061 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3121 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3181 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3241 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3301 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3361 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg
     3421 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     3481 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     3541 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     3601 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     3661 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     3721 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     3781 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     3841 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     3901 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     3961 tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt gagcaagggc
     4021 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     4081 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     4141 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     4201 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     4261 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     4321 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     4381 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     4441 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     4501 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     4561 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     4621 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     4681 cactccaccg gcggcatgga cgagctgtac aagtccggac tcagatctag tggctgtttt
     4741 ccagtttctg gcctccgctg cctatctagg gacggcagga tggccgcgca gggcgcgccg
     4801 cgcttcctcc tgaccttcga cttcgacgag actatcgtgg acgaaaacag cgacgattcg
     4861 atcgtgcgcg ccgcgccggg ccagcggctc ccggagagcc tgcgagccac ctaccgcgag
     4921 ggcttctaca acgagtacat gcagcgcgtc ttcaagtacc tgggcgagca gggcgtgcgg
     4981 ccgcgggacc tgagcgccat ctacgaagcc atccctttgt cgccaggcat gagcgacctg
     5041 ctgcagtttg tggcaaaaca gggcgcctgc ttcgaggtga ttctcatctc cgatgccaac
     5101 acctttggcg tggagagctc gctgcgcgcc gccggccacc acagcctgtt ccgccgcatc
     5161 ctcagcaacc cgtcggggcc ggatgcgcgg ggactgctgg ctctgcggcc gttccacaca
     5221 cacagctgcg cgcgctgccc cgccaacatg tgcaagcaca aggtgctcag cgactacctg
     5281 cgcgagcggg cccacgacgg cgtgcacttc gagcgcctct tctacgtggg cgacggcgcc
     5341 aacgacttct gccccatggg gctgctggcg ggcggcgacg tggccttccc gcgccgcggc
     5401 taccccatgc accgcctcat tcaggaggcc cagaaggccg agcccagctc gttccgcgcc
     5461 agcgtggtgc cctgggaaac ggctgcagat gtgcgcctcc acctgcaaca ggtgctgaag
     5521 tcgtgctga
//
