LOCUS       sequence-258976-        6334 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  nuclear marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6334)
  TITLE     Plasmids used for tagging different cellular components.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6334)
  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            "Unpublished"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: Clim-DC1-mCherry-H2B-Venus
FEATURES             Location/Qualifiers
     source          1..6334
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    53..74
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(89..94,95..112,113..119)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    127..143
                     /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     132..154
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     151..167
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     151..167
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             968..1675
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGAHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1113..1131)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1381..1400
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1575..1598
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             1685..2284
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1685..1704)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2181..2201
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2307..2428
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      3423..3432
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3429..3431,3432..3434,3435..4145)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDRMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3474..3495)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3735..3754)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4082..4103
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4174..4295
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4332..4351
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4480..4548,4549..5340)
                     /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(4698..4717)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5511..6099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6000..6019
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6253..6270
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
       61 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
      121 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
      181 agcttatcgg tgggactgtg agaactgcta cctgcctgtg cagaggagag cacactacat
      241 tcaccattga aaatatggcc cactgaactg atcaagccaa gtgaagactg cgcgcctgtg
      301 ttctgattct tcccatggta caagtactga aactttggaa gcgccagaga attcacagac
      361 gcacagaaga cagcgcatgg tggaggctgt gtgtgttcac cgaggtgtcc tctgcggaag
      421 gtgccgttca ttatgtattc ggagtaaagc acaacctctt caatgctgct cccttccttc
      481 gcttgtttct tggatagtta tcgtctgtca gaatctttcg gtacattcag tatagagttc
      541 gcgatcggta gcaacagtcg gacggaccct gtcaggacgg tgtattcggt ggcctacttt
      601 tcttttgttg catagctgtt cctgacgcga tgcgccaagc gccgtcatat tgtgctgtgt
      661 gcccctatgt ggagcaggca tagagctctt gccaaatgtt gggaagagct gttggagaag
      721 cgacagagca gtgccaagac gacgaacgag gtgtgcgagc tgtagtgcta gtgcttccta
      781 ggttctggtt ttactgacac ctatttgaag gaataatttc tgctcacaca cagttttgta
      841 accgctcact attgtgttac agctcatcat aagctactat aacacatctt tgcggcttta
      901 agctagtgta ttcatctaat ttacaatcat gcgcgagtgt atctctattc ccgggactag
      961 tgatatcatg gtgagcaagg gcgaggagga caacatggcc atcatcaagg agttcatgcg
     1021 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     1081 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc
     1141 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     1201 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     1261 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     1321 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     1381 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     1441 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcgcccacta
     1501 cgacgccgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     1561 caacgtcaac atcaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     1621 gtacgagcgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaaggcggc
     1681 cgccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     1741 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     1801 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     1861 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     1921 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     1981 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     2041 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     2101 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     2161 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     2221 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     2281 ctgacctcga gcatcgatag acatgataag atacattgat gagtttggac aaaccacaac
     2341 aagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2401 aaccattata agctgcaata aacaagttaa caacaacaat tctagagggc ctgcttgccg
     2461 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gcgaaatgct
     2521 gggcgccttg aaactgaacc tgttcggatc cgggttgcgc gcttccattg ttttggtttc
     2581 cgtgtctgca aggcgggtct ggacgtgacg ctttggggga gagggagagc agagatacaa
     2641 ttctagccac gaatttcagc caatagtcac agaggggagt tgcgatcctg cgaattggct
     2701 taggtctgtg aggtctttca cggtttcgaa agcagcgcag aggggagacc gactaccaaa
     2761 agccataaca ctccaccgag atatccgcac tcttcgctag atgagcttgc ttctcctctt
     2821 gtcacaaatc agcccaatga tagtcactgt tcaacaacgg tgcagtgtaa atattcgttg
     2881 tgcgtccttg atgagcagaa ggtacgagaa gcaggaaatc atttcataat ggaaaaccat
     2941 tcactgtctt tctgtattgc gaaaagagtg atcataggta atcgcaacgg cgggtcttta
     3001 gaacagtcac ataccagatc aagcttgtcg acatgccacc tgttgcatcc aagaagggcg
     3061 agaagaaggc cgccggcaag aagggcggag acaagggcgg taagcgcaag aaggcccgca
     3121 aggaatctta ctccatctac atctacaagg ttctcaagca ggtccaccct gacactggtg
     3181 tctccaacaa agccatgttg atcatgaact ccttcgtgtc tgacattttc gagcgtgtcg
     3241 ccaacgaagc ttcccgtctt gccaagtaca acaagaagtc caccatcact tcccgggaga
     3301 ttcagactgc tgtccgtctg ctcctgcctg gtgagcttgc taagcacgcc gtctcagaag
     3361 gtaccaaggc cgtgaccaag tacacttcct ctaagaacgc ggccgcatcc atcacactgg
     3421 cggccgccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     3481 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     3541 ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct
     3601 ggcccaccct cgtgaccacc ctgggctacg gcctgcagtg cttcgcccgc taccccgacc
     3661 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     3721 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     3781 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     3841 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc
     3901 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
     3961 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     4021 acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     4081 gcatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     4141 acaagctcga gtaagctagc agatctaggg ccctaagata cattgatgag tttggacaaa
     4201 ccacaacaag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     4261 tatttgtaac cattataagc tgcaataaac aagttgccgg ctgggtgtgg cggaccgcta
     4321 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4381 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4441 ccttcttgac gagttcttct gaattgaaaa aggaagagta tgagtattca acatttccgt
     4501 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4561 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4621 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4681 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4741 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4801 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4861 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4921 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4981 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5041 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5101 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5161 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5221 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5281 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5341 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5401 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5461 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5521 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5581 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5641 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5701 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5761 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5821 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5881 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5941 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6001 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6061 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6121 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6181 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6241 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     6301 cctctccccg cgcgttggcc gattcattaa tgca
//
