LOCUS       sequence-271958-        6858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral plasmid expressing mCherry - HJURP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6858)
  TITLE     Cheeseman Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6858)
  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            "Unpublished"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: pKM510
FEATURES             Location/Qualifiers
     source          1..6858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        81..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..588
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     539..559
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      627..636
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             633..1340
                     /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(778..796)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1046..1065
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1386..1406
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1419..1463
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2452..2471)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3513..3533
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3666..3787
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3703..3722)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3757..3776
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3794..4249)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3931..3952)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4143..4162
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4276..4380
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4382..4739
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4382..4573
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4405..4425)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4590..4725
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4774..5568
                     /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             4774..5568
                     /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(4828..4847)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5438..5457
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5756..5775)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5800..5847
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6176..6764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6665..6684
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      421 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      481 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      541 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
      601 cgtcagatcc gctagcgcta ccggtcgcca ccatggtgag caagggcgag gaggataaca
      661 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
      721 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
      781 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
      841 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
      901 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
      961 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     1021 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     1081 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
     1141 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
     1201 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
     1261 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     1321 gcatggacga gctgtacaag tccggagggg atcggtggag ctccaccggt ggcggccgct
     1381 ctagagagaa tctttatttt cagggcgccg ccaaattcaa agaaaccgct gctgctaaat
     1441 tcgaacgcca gcacatggac agcggaggtg gaggtactag tctcgagatg ctgggtacgc
     1501 tgcgcgccat ggagggcgag gacgtggaag acgaccagct gctgcagaag ctcagggcca
     1561 gtcgccgccg cttccagagg cgcatgcagc ggctgataga gaagtacaac cagcccttcg
     1621 aggacacccc ggtggtgcaa atggccacgc tgacctacga gacgccacag ggattgagaa
     1681 tttggggtgg aagactaata aaggaaagaa acaaaggaga gatccaggac tcctccatga
     1741 agcccgcgga caggacagat ggctccgtgc aagctgcagc ctggggtcct gagcttccct
     1801 cgcaccgcac agtcctggga gccgattcaa aaagcggtga ggtcgatgcc acgtcagacc
     1861 aggaagagtc agttgcttgg gccttagcac ctgcagtgcc tcaaagccct ttgaaaaatg
     1921 aattaagaag gaaatacttg acccaagtgg atatactgct acaaggtgca gagtattttg
     1981 agtgtgcagg taacagagct ggaagggatg tacgtgtgac tccgctgcct tcactggcct
     2041 cacctgccgt gcctgccccc ggatactgca gtcgtatctc cggaaagagt cctggtgacc
     2101 cagcgaaacc agcttcatct cccagagaat gggatccttt gcatccttcc tccacagaca
     2161 tggccttagt acctagaaat gacagcctct ccctacaaga gaccagtagc agcagcttct
     2221 taagcagcca gccctttgaa gatgatgaca tttgcaatgt gaccatcagt gacctgtacg
     2281 cagggatgct gcactccatg agccggctgt tgagcacaaa gccatcaagc atcatctcca
     2341 ccaaaacgtt catcatgcaa aactggaact gcaggaggag gcacagatat aagagcaggg
     2401 aattcccgcg gggaagtgga agtggatcca tggtgaaacc agtaacgtta tacgatgtcg
     2461 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg
     2521 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca
     2581 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca
     2641 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac
     2701 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg
     2761 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg
     2821 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg
     2881 tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg
     2941 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa
     3001 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa
     3061 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaaccr
     3121 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg
     3181 cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg
     3241 tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca
     3301 aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg
     3361 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc
     3421 tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     3481 cacgacaggt ttcccgaagc agcctgaggc ctcctaagaa gaagaggaag gtttgaggat
     3541 ccaccggatc tagataactg atcataatca gccataccac atttgtagag gttttacttg
     3601 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3661 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3721 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3781 tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3841 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3901 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3961 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     4021 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     4081 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     4141 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     4201 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     4261 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4321 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4381 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4441 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4501 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4561 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4621 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     4681 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     4741 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4801 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4861 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4921 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     4981 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5041 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5101 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5161 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5221 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5281 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     5341 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5401 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5461 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5521 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5581 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5641 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5701 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     5761 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     5821 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     5881 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     5941 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     6001 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     6061 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6121 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6181 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6241 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6301 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6361 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6421 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6481 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6541 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6601 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6661 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6721 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6781 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6841 cccctgattc tgtggata
//
