LOCUS       sequence-258977-        4721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cytoplasmic marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4721)
  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 4721)
  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: CTMP
FEATURES             Location/Qualifiers
     source          1..4721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    32..53
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(68..73,74..91,92..98)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    106..122
                     /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     111..133
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     130..146
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     130..146
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             947..1654
                     /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(1092..1110)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1360..1379
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1554..1577
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             1664..2263
                     /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(1664..1683)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2160..2180
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2698..2717
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2846..2914,2915..3706)
                     /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(3064..3083)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3877..4465
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4366..4385
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4619..4636
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
       61 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
      121 aatttcacac aggaaacagc tatgaccatg attacgccaa gcttatcggt gggactgtga
      181 gaactgctac ctgcctgtgc agaggagagc acactacatt caccattgaa aatatggccc
      241 actgaactga tcaagccaag tgaagactgc gcgcctgtgt tctgattctt cccatggtac
      301 aagtactgaa actttggaag cgccagagaa ttcacagacg cacagaagac agcgcatggt
      361 ggaggctgtg tgtgttcacc gaggtgtcct ctgcggaagg tgccgttcat tatgtattcg
      421 gagtaaagca caacctcttc aatgctgctc ccttccttcg cttgtttctt ggatagttat
      481 cgtctgtcag aatctttcgg tacattcagt atagagttcg cgatcggtag caacagtcgg
      541 acggaccctg tcaggacggt gtattcggtg gcctactttt cttttgttgc atagctgttc
      601 ctgacgcgat gcgccaagcg ccgtcatatt gtgctgtgtg cccctatgtg gagcaggcat
      661 agagctcttg ccaaatgttg ggaagagctg ttggagaagc gacagagcag tgccaagacg
      721 acgaacgagg tgtgcgagct gtagtgctag tgcttcctag gttctggttt tactgacacc
      781 tatttgaagg aataatttct gctcacacac agttttgtaa ccgctcacta ttgtgttaca
      841 gctcatcata agctactata acacatcttt gcggctttaa gctagtgtat tcatctaatt
      901 tacaatcatg cgcgagtgta tctctattcc cgggactagt gatatcatgg tgagcaaggg
      961 cgaggaggac aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     1021 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     1081 cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat
     1141 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     1201 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     1261 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     1321 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     1381 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     1441 gatcaagcag aggctgaagc tgaaggacgg cgcccactac gacgccgagg tcaagaccac
     1501 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagctgga
     1561 catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgcg ccgagggccg
     1621 ccactccacc ggcggcatgg acgagctgta caaggcggcc gccatgaccg agtacaagcc
     1681 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     1741 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     1801 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     1861 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     1921 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     1981 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     2041 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     2101 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     2161 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     2221 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacctcgag taaacaaatg
     2281 ttttgttgga agatttgatt tgtctttttt ggagtgctta gttagtgcac tcctagatga
     2341 gcaaacatta ccacagtgta tcgcagcata aaataaaatt ataaaatgtc tttgtagatc
     2401 aaaagcatta caaacaaata tttaaacttt gatcactttt ttcaaattcg tcaatatgac
     2461 taattaacat ctgggttata gatgacaagc atgaatgttt catttatttt aaaaaaaaaa
     2521 taaataagta ttttttcatt tgcatcacca tcaaaaatat ttccattgaa aagtcatcga
     2581 ctggctattt aaatttctag agggcctgct tgccgaatat catggtggaa aatggccgct
     2641 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     2701 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     2761 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     2821 tcttctgaat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     2881 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     2941 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3001 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3061 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3121 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3181 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3241 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3301 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3361 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     3421 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     3481 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     3541 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     3601 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     3661 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     3721 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3781 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3841 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3901 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3961 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4021 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4081 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4141 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4201 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4261 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4321 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4381 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4441 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4501 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     4561 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     4621 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     4681 ttggccgatt cattaatgca gctggcacga caggtttccc g
//
