LOCUS       sequence-268636-        4999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mMoClo TUPV, with ZF43x1 promoter and mKate2 in the MCS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4999)
  AUTHORS   Donahue PS, Draut JW, Muldoon JJ, Edelstein HI, Bagheri N, Leonard 
            JN
  TITLE     The COMET toolkit for composing customizable genetic programs in 
            mammalian cells.
  JOURNAL   Nat Commun. 2020 Feb 7;11(1):779. doi: 10.1038/s41467-019-14147-5.
  PUBMED    32034124
REFERENCE   2  (bases 1 to 4999)
  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: "Nat 
            Commun."; date: "2020-02-7"; volume: "11(1):779. doi"; pages: " 
            10.1038/s41467-019-14147-5"
COMMENT     Sequence Label: pHIE043 ZF43x1 mKate2 (TUPV1)
FEATURES             Location/Qualifiers
     source          1..4999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      160..169
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             166..864
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     primer_bind     complement(909..926)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    915..1139
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1908..2336
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1995..2014)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2205..2226
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2359..2555
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2406..2541
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2468..2487
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2675..2796
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2712..2731)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2766..2785
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2845..2861)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2845..2861)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2858..2880)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2869..2885
                     /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)."
     promoter        complement(join(2893..2899,2900..2917,2918..2923))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2938..2959
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3076..3093)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3247..3832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3327..3346)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4003..4794,4795..4863))
                     /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     4626..4645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4864..4968)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgaag acggtgccaa ggtcttgaca aacgagtgct agtacaagat
       61 cgcgtcgacg gatcgggaga tctgaattag agtgaggacg atccctctag agggtatata
      121 atgggggcca ctagtctact accagagctc atcgctagcg ccaccatggt gagcgagctg
      181 attaaggaga acatgcacat gaagctgtat atggagggca ccgtgaacaa ccaccacttc
      241 aagtgtacct ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag
      301 gcggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctaccag cttcatgtac
      361 ggcagcaaaa ccttcatcaa ccacacccag ggcatccccg acttctttaa gcagtccttc
      421 cccgagggct tcacatggga gagagtcacc acatacgaag atgggggcgt gctgaccgct
      481 acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg
      541 aacttcccat ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggcctccacc
      601 gaaaccctgt accccgctga cggcggcctg gaaggcagag ccgacatggc cctgaagctc
      661 gtgggcgggg gccacctgat ctgcaacttg aaaaccacat acagatccaa gaaacccgct
      721 aagaacctca agatgcccgg cgtctactat gtggacagga gactggaaag aatcaaggag
      781 gccgacaaag aaacctacgt cgagcagcac gaggtggctg tggccagata ctgcgacctc
      841 cctagcaaac tggggcacag atgagcggcc gctcgagtct agagggcccg tttaaacccg
      901 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
      961 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     1021 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     1081 caagggggag gattgggaag tcaatagcag gcatgctggg gatgcggtgg gctctatggc
     1141 ttctgaggca ggtcgagggt acatgcgcct tactccttgt gagcgcatcg cgataagctc
     1201 tagagggaca gccccccccc aaagccccca gggatgtaat tacgtccctc ccccgctagg
     1261 gggcagcagc gagccgcccg gggctccgct ccggtccggc gctccccccg catccccgag
     1321 ccggcagcgt gcggggacag cccgggcacg gggaaggtgg cacgggatcg ctttcctctg
     1381 aacgcttctc gctgctcttt gagcctgcag acacctgggg ggatacgggg aaaaagcttt
     1441 aggctgaaag agagatttag aatgacagaa tcatagaacg gcctgcgtac ggatggggct
     1501 ctagctctag agcttgatat cgaattcctg cagccccgct agagggacag ccccccccca
     1561 aagcccccag ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg
     1621 ggctccgctc cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc
     1681 ccgggcacgg ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg
     1741 agcctgcaga cacctggggg gatacgggga aaaagcttta ggctgaaaga gagatttaga
     1801 atgacagaat catagaacgg cctgggcgcg cctggtggcg aacgatcaac agacgggaga
     1861 gggcaaaagt cttcgctgac agcgaccgga ctctaggggg tatccccacg cgccctgtag
     1921 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1981 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2041 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2101 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2161 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2221 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2281 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     2341 ctgtggaatg tgtgtcagtt aggctcaatt agtcagcaac catagtcccg cccctaactc
     2401 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2461 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     2521 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     2581 ttttcggagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     2641 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     2701 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2761 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     2821 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2881 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2941 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3001 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3061 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3121 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3181 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3241 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3301 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3361 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3421 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3481 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3541 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3601 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3661 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3721 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3781 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3841 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3901 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3961 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4021 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4081 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4141 gcgagatcca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4201 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4261 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4321 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4381 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4441 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4501 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4561 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4621 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4681 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4741 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4801 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4861 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4921 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4981 aaaagtgcca cctgacgtc
//
