LOCUS       sequence-271954-        8920 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 8920)
  TITLE     Cheeseman Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8920)
  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            "Unpublished"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: pKM28
FEATURES             Location/Qualifiers
     source          1..8920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        127..430
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        431..634
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     585..605
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      673..682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(679..681,682..684,685..1395)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(679..681,682..684,685..1395)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(724..745)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(985..1004)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1332..1353
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1441..1461
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1474..1518
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3839..3858)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4900..4920
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          5083..5148
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    5308..5429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    5774..5895
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(5902..6357)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6039..6060)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6251..6270
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6384..6488
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6490..6847
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6490..6681
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6513..6533)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6698..6833
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6882..7676
                     /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             6882..7676
                     /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(6936..6955)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7546..7565
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7864..7883)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7908..7955
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8284..8872
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8773..8792
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
       61 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      121 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      181 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      241 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      301 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      361 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      421 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      481 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      541 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      601 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      661 gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      721 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      781 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      841 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
      901 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      961 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     1021 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     1081 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     1141 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     1201 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1261 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     1321 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1381 gacgagctgt acaagtccgg aggggatcgg tggagctcca ccggtggcgg ccgctctaga
     1441 gagaatcttt attttcaggg cgccgccaaa ttcaaagaaa ccgctgctgc taaattcgaa
     1501 cgccagcaca tggacagcgg aggtggaggt actagtctcg agatgctggg tacgctgcgc
     1561 gccatggagg gcgaggacgt ggaagacgac cagctgctgc agaagctcag ggccagtcgc
     1621 cgccgcttcc agaggcgcat gcagcggctg atagagaagt acaaccagcc cttcgaggac
     1681 accccggtgg tgcaaatggc cacgctgacc tacgagacgc cacagggatt gagaatttgg
     1741 ggtggaagac taataaagga aagaaacaaa ggagagatcc aggactcctc catgaagccc
     1801 gcggacagga cagatggctc cgtgcaagct gcagcctggg gtcctgagct tccctcgcac
     1861 cgcacagtcc tgggagccga ttcaaaaagc ggtgaggtcg atgccacgtc agaccaggaa
     1921 gagtcagttg cttgggcctt agcacctgca gtgcctcaaa gccctttgaa aaatgaatta
     1981 agaaggaaat acttgaccca agtggatata ctgctacaag gtgcagagta ttttgagtgt
     2041 gcaggtaaca gagctggaag ggatgtacgt gtgactccgc tgccttcact ggcctcacct
     2101 gccgtgcctg cccccggata ctgcagtcgt atctccggaa agagtcctgg tgacccagcg
     2161 aaaccagctt catctcccag agaatgggat cctttgcatc cttcctccac agacatggcc
     2221 ttagtaccta gaaatgacag cctctcccta caagagacca gtagcagcag cttcttaagc
     2281 agccagccct ttgaagatga tgacatttgc aatgtgacca tcagtgacct gtacgcaggg
     2341 atgctgcact ccatgagccg gctgttgagc acaaagccat caagcatcat ctccaccaaa
     2401 acgttcatca tgcaaaactg gaactgcagg aggaggcaca gatataagag caggatgaac
     2461 aaaacatatt gcaaaggagc cagacgttct cagaggagct ccaaggagaa cttcataccc
     2521 tgctctgagc ctgtgaaagg gacaggggca ttaagagatt gcaagaacgt attagatgtt
     2581 tcttgccgta agacaggttt aaaattggaa aaagcttttc ttgaagtcaa cagaccccaa
     2641 atccataagt tagatccaag ttggaaggag cgcaaagtga caccctcgaa gtattcttcc
     2701 ttgatttact tcgactccag tgcaacatat aatcttgatg aggaaaatag atttaggaca
     2761 ttaaaatggt taatttctcc tgtaaaaata gtttccagac caacaatacg acagggccat
     2821 ggagagaacc gtcagaggga gattgaaatc cgatttgatc agcttcatcg ggaatattgc
     2881 ctgagtccca ggaaccagcc tcgccggatg tgcctcccgg actcctgggc catgaacatg
     2941 tacagagggg gtcctgcgag tcctggtggc cttcagggct tagaaacccg caggctgagt
     3001 ttaccttcca gcaaagcaaa agcaaaaagt ttaagtgagg cttttgaaaa cctaggcaaa
     3061 agatctctgg aagcaggtag gtgcctgccc aagagcgatt catcttcatc acttccaaag
     3121 accaacccca cacacagcgc aactcgcccg cagcagacat ctgaccttca cgttcaggga
     3181 aatagttctg gaatatttag aaagtcagtg tcacccagca aaactctttc agtcccagat
     3241 aaagaagtgc caggccacgg aaggaatcgt tacgatgaaa ttaaagaaga atttgacaag
     3301 cttcatcaaa agtattgcct caaatctcct gggcagatga cagtgccttt atgtattgga
     3361 gtgtctacag ataaagcaag tatggaagtt cgatatcaaa cagaaggctt cttaggaaaa
     3421 ttaaatccag accctcactt ccagggtttc cagaagttgc catcatcacc cctggggtgc
     3481 agaaaaagtc tactgggctc aactgcaatt gaggctcctt catctacatg tgttgctcgt
     3541 gccatcacga gggatggcac gagggaccat cagttccctg caaaaagacc caggctatca
     3601 gaaccccagg gctccggacg ccagggcaat tccctgggtg cctcagatgg ggtggacaac
     3661 accgtcagac cgggagacca gggcagctct tcacagccca actcagaaga gagaggagag
     3721 aacacgtctt acaggatgga agagaaaagt gatttcatgc tagaaaaatt ggaaactaaa
     3781 agtgtggaat tcccgcgggg aagtggaagt ggatccatgg tgaaaccagt aacgttatac
     3841 gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc
     3901 agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac
     3961 attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc
     4021 acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc
     4081 gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt
     4141 aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg
     4201 ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt
     4261 cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg
     4321 cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc
     4381 ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc
     4441 aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa
     4501 caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat
     4561 cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat
     4621 atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc gccgttaacc
     4681 accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc
     4741 tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa
     4801 accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     4861 cagctggcac gacaggtttc ccgaagcagc ctgaggcctc ctaagaagaa gaggaaggtt
     4921 tgagcgcaac gcaattaatg taagttagct cactcattag gcaccccagg ctttacactt
     4981 tatgcttccg acctgcagcc ccttgggatc tttgtgaagg aaccttactt ctgtggtgtg
     5041 acataattgg acaaactacc tacagagatt taaagctcta aggtaaatat aaaattttta
     5101 agtgtataat gtgttaaact actgattcta attgtttgtg tattttagat tcacagtccc
     5161 aaggctcatt tcaggcccct cagtcctcac agtctgttca tgatcataat cagccatacc
     5221 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     5281 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     5341 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5401 ggtttgtcca aactcatcaa tgtatcttat catgtctggg cccatcgatg gcactgggat
     5461 ctgcgaacgc cagcaagacg tagcccagcg cgtcggcccc gagatgcgcc gcgtgcggct
     5521 gctggagatg gcggacgcga tggatatgtt ctgccaaggg ttggtttgcg cattcacagt
     5581 tctccgcaag aattgattgg ctccaattct tggagtggtg aatccgttag cgaggtgccg
     5641 ccctgcttca tccccgtggc ccgttgctcg cgtttgctgg cggtgtcccc ggaagaaata
     5701 tatttgcatg tctttagttc tatgatgaca caaaccccgc ccagcgtctt gtcattggcg
     5761 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5821 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5881 catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     5941 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     6001 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     6061 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     6121 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     6181 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     6241 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     6301 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     6361 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     6421 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     6481 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     6541 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     6601 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     6661 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     6721 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     6781 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     6841 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     6901 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     6961 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     7021 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     7081 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     7141 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     7201 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     7261 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     7321 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     7381 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     7441 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     7501 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     7561 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     7621 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     7681 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     7741 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     7801 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     7861 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     7921 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     7981 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     8041 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     8101 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     8161 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     8221 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     8281 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     8341 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     8401 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     8461 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     8521 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     8581 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     8641 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     8701 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     8761 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     8821 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     8881 caacgcggcc tttttacggt tcctggcctt ttgctggcct
//
