LOCUS       sequence-258968-        5962 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pClimDC-NMM:tdT cell membrane marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5962)
  AUTHORS   Kozyczkowska A, Najle SR, Ocana-Pallares E, Areste C, Shabardina V, 
            Ara PS, Ruiz-Trillo I, Casacuberta E
  TITLE     Stable transfection in protist Corallochytrium limacisporum 
            identifies novel cellular features among unicellular animals 
            relatives.
  JOURNAL   Curr Biol. 2021 Sep 27;31(18):4104-4110.e5. doi: 
            10.1016/j.cub.2021.06.061. Epub 2021 Jul 21.
  PUBMED    34293333
REFERENCE   2  (bases 1 to 5962)
  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"; doi: 
            "10.1016/j.cub.2021.06.061"; journalName: "Curr Biol"; date: 
            "2021-09-27- 27"; volume: "31"; issue: "18"; pages: "4104-4110.e5"
COMMENT     Sequence Label: Clim-DC2-NMM-tdtomato
FEATURES             Location/Qualifiers
     source          1..5962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     13..30
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    147..168
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(183..188,189..206,207..213)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    221..237
                     /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     226..248
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     245..261
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     245..261
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1050..1649
                     /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(1050..1069)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1546..1566
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    1672..1793
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             2444..3871
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYE
                     GTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMN
                     FEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVL
                     KGEIHQALKLKDGGRYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYER
                     SEGRHHLFLYGMDELYK"
     polyA_signal    3896..4017
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4054..4073
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4202..4270,4271..5062)
                     /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(4420..4439)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5233..5821
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5722..5741
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
       61 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
      121 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
      181 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
      241 cacacaggaa acagctatga ccatgattac gccaagctta tcggtgggac tgtgagaact
      301 gctacctgcc tgtgcagagg agagcacact acattcacca ttgaaaatat ggcccactga
      361 actgatcaag ccaagtgaag actgcgcgcc tgtgttctga ttcttcccat ggtacaagta
      421 ctgaaacttt ggaagcgcca gagaattcac agacgcacag aagacagcgc atggtggagg
      481 ctgtgtgtgt tcaccgaggt gtcctctgcg gaaggtgccg ttcattatgt attcggagta
      541 aagcacaacc tcttcaatgc tgctcccttc cttcgcttgt ttcttggata gttatcgtct
      601 gtcagaatct ttcggtacat tcagtataga gttcgcgatc ggtagcaaca gtcggacgga
      661 ccctgtcagg acggtgtatt cggtggccta cttttctttt gttgcatagc tgttcctgac
      721 gcgatgcgcc aagcgccgtc atattgtgct gtgtgcccct atgtggagca ggcatagagc
      781 tcttgccaaa tgttgggaag agctgttgga gaagcgacag agcagtgcca agacgacgaa
      841 cgaggtgtgc gagctgtagt gctagtgctt cctaggttct ggttttactg acacctattt
      901 gaaggaataa tttctgctca cacacagttt tgtaaccgct cactattgtg ttacagctca
      961 tcataagcta ctataacaca tctttgcggc tttaagctag tgtattcatc taatttacaa
     1021 tcatgcgcga gtgtatctct attcccggga tgaccgagta caagcccacg gtgcgcctcg
     1081 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     1141 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     1201 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     1261 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     1321 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     1381 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     1441 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     1501 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     1561 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     1621 ggtgcatgac ccgcaagccc ggtgcctgac ctcgagcatc gatagacatg ataagataca
     1681 ttgatgagtt tggacaaacc acaacaagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     1741 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     1801 acaattctag agggcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     1861 catcgactgt ggccggcgaa atgctgggcg ccttgaaact gaacctgttc ggatccgggt
     1921 tgcgcgcttc cattgttttg gtttccgtgt ctgcaaggcg ggtctggacg tgacgctttg
     1981 ggggagaggg agagcagaga tacaattcta gccacgaatt tcagccaata gtcacagagg
     2041 ggagttgcga tcctgcgaat tggcttaggt ctgtgaggtc tttcacggtt tcgaaagcag
     2101 cgcagagggg agaccgacta ccaaaagcca taacactcca ccgagatatc cgcactcttc
     2161 gctagatgag cttgcttctc ctcttgtcac aaatcagccc aatgatagtc actgttcaac
     2221 aacggtgcag tgtaaatatt cgttgtgcgt ccttgatgag cagaaggtac gagaagcagg
     2281 aaatcatttc ataatggaaa accattcact gtctttctgt attgcgaaaa gagtgatcat
     2341 aggtaatcgc aacggcgggt ctttagaaca gtcacatacc agatcaagct tgatgggcgg
     2401 ttgctgtagt acggagactc aacagcggca gcagcctgta cgggttagca agggcgagga
     2461 ggtcatcaaa gagttcatgc gcttcaaggt gcgcatggag ggctccatga acggccacga
     2521 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     2581 gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcat
     2641 gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc cccgattaca agaagctgtc
     2701 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gtctggtgac
     2761 cgtgacccag gactcctccc tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg
     2821 caccaacttc ccccccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     2881 caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccacc aggccctgaa
     2941 gctgaaggac ggcggccact acctggtgga gttcaagacc atctacatgg ccaagaagcc
     3001 cgtgcaactg cccggctact actacgtgga caccaagctg gacatcacct cccacaacga
     3061 ggactacacc atcgtggaac agtacgagcg ctccgagggc cgccaccacc tgttcctggg
     3121 gcatggcacc ggcagcaccg gcagcggcag ctccggcacc gcctcctccg aggacaacaa
     3181 catggccgtc atcaaagagt tcatgcgctt caaggtgcgc atggagggct ccatgaacgg
     3241 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     3301 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca
     3361 gttcatgtac ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg attacaagaa
     3421 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggtct
     3481 ggtgaccgtg acccaggact cctccctgca ggacggcacg ctgatctaca aggtgaagat
     3541 gcgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacca tgggctggga
     3601 ggcctccacc gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccaccaggc
     3661 cctgaagctg aaggacggcg gccgctacct ggtggagttc aagaccatct acatggccaa
     3721 gaagcccgtg caactgcccg gctactacta cgtggacacc aagctggaca tcacctccca
     3781 caacgaggac tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctgtt
     3841 cctgtacggc atggacgagc tgtacaagtg acatatgcta gcagatctag ggccctaaga
     3901 tacattgatg agtttggaca aaccacaaca agaatgcagt gaaaaaaatg ctttatttgt
     3961 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttgcc
     4021 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4081 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4141 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgaattgaa aaaggaagag
     4201 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4261 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4321 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4381 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4441 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4501 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4561 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4621 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4681 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4741 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     4801 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4861 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4921 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4981 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5041 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5101 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5161 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5221 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5281 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5341 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5401 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5461 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5521 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5581 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5641 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5701 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5761 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5821 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5881 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     5941 tgataccgct cgccgcagcc ga
//
