LOCUS       sequence-272475-        5004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of EB3-tDT BMAM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5004)
  TITLE     Cheeseman Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5004)
  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            "Unpublished"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: pDK712
FEATURES             Location/Qualifiers
     source          1..5004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(75..108)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        186..565
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        566..769
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     720..740
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1759..1768
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1765..2466
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(1898..1916)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2166..2185
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2360..2383
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2635..2769
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2672..2691)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2726..2745
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2890..3055
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3245..3833
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3734..3753
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  3968..4191
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4258..4791)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4980..4985,4986..5002,5003..5004,1..4))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 tcaactgggt tcgtgccttc atccgtttcc acggtgtgcg tcacccggca accttgggca
       61 gcagcgaagt cgccataact tcgtatagca tacattatac gaagttatct gtaactataa
      121 cggtcctaag gtagcgagtt taaacactag tatcgatcgc gatgtacggg ccagatatac
      181 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      241 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      301 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      361 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      421 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      481 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      541 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      601 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      661 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      721 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
      781 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gccggcaaat
      841 tctcgagtag gatcccggtc cgaagcgcgc ggaattcaaa ggcctacgtc gactgccacc
      901 atggccgtca atgtgtactc cacatctgtg accagtgaaa atctgagtcg ccatgatatg
      961 cttgcatggg tcaacgactc cctgcacctc aactatacca agatagaaca gctttgttca
     1021 ggggcagcct actgccagtt catggacatg ctcttccccg gctgtgtgca cttgaggaaa
     1081 gtgaagttcc aggccaaact agagcatgaa tacatccaca acttcaaggt gctgcaagca
     1141 gctttcaaga agatgggtgt tgacaaaatc attcctgtag agaaattagt gaaaggaaaa
     1201 ttccaagata attttgagtt tattcagtgg tttaagaaat tctttgacgc aaactatgat
     1261 ggaaaggatt acaaccctct gctggcgcgg cagggccagg acgtagcgcc acctcctaac
     1321 ccaggtgatc agatcttcaa caaatccaag aaactcattg gcacagcagt tccacagagg
     1381 acgtccccca caggcccaaa aaacatgcag acctctggcc ggctgagcaa tgtggccccc
     1441 ccctgcattc tccggaagaa tcctccatca gcccgaaatg gcggccatga gactgatgcc
     1501 caaattcttg aactcaacca acagctggtg gacttgaagc tgacagtgga tgggctggag
     1561 aaggaacgtg acttctactt cagcaaactt cgtgacatcg agctcatctg ccaggagcat
     1621 gaaagtgaaa acagccctgt tatctcaggc atcattggca tcctctatgc cacagaggaa
     1681 ggattcgcac cccctgagga cgatgagatt gaagagcatc aacaagaaga ccaggacgag
     1741 tacctggatc caccggtcgc caccatggtg agcaagggcg aggaggtcat caaagagttc
     1801 atgcgcttca aggtgcgcat ggagggctcc atgaacggcc acgagttcga gatcgagggc
     1861 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc
     1921 ggccccctgc ccttcgcctg ggacatcctg tccccccagt tcatgtacgg ctccaaggcg
     1981 tacgtgaagc accccgccga catccccgat tacaagaagc tgtccttccc cgagggcttc
     2041 aagtgggagc gcgtgatgaa cttcgaggac ggcggtctgg tgaccgtgac ccaggactcc
     2101 tccctgcagg acggcacgct gatctacaag gtgaagatgc gcggcaccaa cttccccccc
     2161 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac
     2221 ccccgcgacg gcgtgctgaa gggcgagatc caccaggccc tgaagctgaa ggacggcggc
     2281 cactacctgg tggagttcaa gaccatctac atggccaaga agcccgtgca actgcccggc
     2341 tactactacg tggacaccaa gctggacatc acctcccaca acgaggacta caccatcgtg
     2401 gaacagtacg agcgctccga gggccgccac cacctgttcc tgtacggcat ggacgagctg
     2461 tacaagtagg cggccgcttt cgaatctaga gcctgcagtc tcgacaagct tgtcgagaag
     2521 tactagagga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2581 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     2641 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2701 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     2761 gtctggatct gatcactgct tgagcctaga agatccggct gctaacaaag cccgaaagga
     2821 agctgagttg gctgctgcca ccgctgagca ataactatca taacccctag ggtataccca
     2881 tctaattgga accagataag tgaaatctag ttccaaacta ttttgtcatt tttaattttc
     2941 gtattagctt acgacgctac acccagttcc catctatttt gtcactcttc cctaaataat
     3001 ccttaaaaac tccatttcca cccctcccag ttcccaacta ttttgtccgc ccacaaccgg
     3061 ttgacttggg tcaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     3121 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3181 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3241 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3301 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3361 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3421 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3481 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3541 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3601 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3661 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3721 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3781 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3841 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     3901 ctgcgttatc ccctgattga cttgggtcgc tcttcctgtg gatgcgcaga tgccctgcgt
     3961 aagcgggtgt gggcggacaa taaagtctta aactgaacaa aatagatcta aactatgaca
     4021 ataaagtctt aaactagaca gaatagttgt aaactgaaat cagtccagtt atgctgtgaa
     4081 aaagcatact ggacttttgt tatggctaaa gcaaactctt cattttctga agtgcaaatt
     4141 gcccgtcgta ttaaagaggg gcgtggccaa gggcatgtaa agactatatt cgcggcgttg
     4201 tgacaattta ccgaacaact ccgcggccgg gaagccgatc tcggcttgaa cgaattgtta
     4261 ggtggcggta cttgggtcga tatcaaagtg catcacttct tcccgtatgc ccaactttgt
     4321 atagagagcc actgcgggat cgtcaccgta atctgcttgc acgtagatca cataagcacc
     4381 aagcgcgttg gcctcatgct tgaggagatt gatgagcgcg gtggcaatgc cctgcctccg
     4441 gtgctcgccg gagactgcga gatcatagat atagatctca ctacgcggct gctcaaactt
     4501 gggcagaacg taagccgcga gagcgccaac aaccgcttct tggtcgaagg cagcaagcgc
     4561 gatgaatgtc ttactacgga gcaagttccc gaggtaatcg gagtccggct gatgttggga
     4621 gtaggtggct acgtctccga actcacgacc gaaaagatca agagcagccc gcatggattt
     4681 gacttggtca gggccgagcc tacatgtgcg aatgatgccc atacttgagc cacctaactt
     4741 tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctgcgtaaca tcgttgctgc
     4801 tccataacat caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca
     4861 tagactgtac aaaaaaacag tcataacaag ccatgaaaac cgccactgcg ccgttaccac
     4921 cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac ttgcattaca
     4981 gtttacgaac cgaacaggct tatg
//
