LOCUS       sequence-317934-        4987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mMoClo TUPV, with ZF43x0 promoter and mKate2 in the MCS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4987)
  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 4987)
  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: pHIE042 ZF43x0 mKate2 (TUPV1)
FEATURES             Location/Qualifiers
     source          1..4987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      281..290
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             287..985
                     /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(1030..1047)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1036..1260
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2029..2457
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2116..2135)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2326..2347
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2480..2676
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2527..2662
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2589..2608
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2796..2917
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2833..2852)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2887..2906
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2966..2982)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2966..2982)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2979..3001)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2990..3006
                     /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(3014..3020,3021..3038,3039..3044))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3059..3080
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3197..3214)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3368..3956)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3448..3467)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4127..4918,4919..4987))
                     /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     4750..4769
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct gacgtcgacg gatcgggaga tcgaagacgg tgccaaggtc ttgacaaacg
      181 agtgctagta caagatctga attcgagacc acaggtctcg gatccctcta gagggtatat
      241 aatgggggcc actagtctac taccagagct catcgctagc gccaccatgg tgagcgagct
      301 gattaaggag aacatgcaca tgaagctgta tatggagggc accgtgaaca accaccactt
      361 caagtgtacc tccgagggcg aaggcaagcc ctacgagggc acccagacca tgagaatcaa
      421 ggcggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacca gcttcatgta
      481 cggcagcaaa accttcatca accacaccca gggcatcccc gacttcttta agcagtcctt
      541 ccccgagggc ttcacatggg agagagtcac cacatacgaa gatgggggcg tgctgaccgc
      601 tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga tcagaggggt
      661 gaacttccca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg aggcctccac
      721 cgaaaccctg taccccgctg acggcggcct ggaaggcaga gccgacatgg ccctgaagct
      781 cgtgggcggg ggccacctga tctgcaactt gaaaaccaca tacagatcca agaaacccgc
      841 taagaacctc aagatgcccg gcgtctacta tgtggacagg agactggaaa gaatcaagga
      901 ggccgacaaa gaaacctacg tcgagcagca cgaggtggct gtggccagat actgcgacct
      961 ccctagcaaa ctggggcaca gatgagcggc cgctcgagtc tagagggccc gtttaaaccc
     1021 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     1081 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     1141 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     1201 gcaaggggga ggattgggaa gtcaatagca ggcatgctgg ggatgcggtg ggctctatgg
     1261 cttctgaggc aggtcgaggg tacatgcgcc ttactccttg tgagcgcatc gcgataagct
     1321 ctagagggac agcccccccc caaagccccc agggatgtaa ttacgtccct cccccgctag
     1381 ggggcagcag cgagccgccc ggggctccgc tccggtccgg cgctcccccc gcatccccga
     1441 gccggcagcg tgcggggaca gcccgggcac ggggaaggtg gcacgggatc gctttcctct
     1501 gaacgcttct cgctgctctt tgagcctgca gacacctggg gggatacggg gaaaaagctt
     1561 taggctgaaa gagagattta gaatgacaga atcatagaac ggcctgcgta cggatggggc
     1621 tctagctcta gagcttgata tcgaattcct gcagccccgc tagagggaca gccccccccc
     1681 aaagccccca gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg
     1741 gggctccgct ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag
     1801 cccgggcacg gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt
     1861 gagcctgcag acacctgggg ggatacgggg aaaaagcttt aggctgaaag agagatttag
     1921 aatgacagaa tcatagaacg gcctgggcgc gcctggtggc gaacgatcaa cagacgggag
     1981 agggcaaaag tcttcgctga cagcgaccgg actctagggg gtatccccac gcgccctgta
     2041 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2101 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2161 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2221 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2281 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2341 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     2401 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     2461 tctgtggaat gtgtgtcagt taggctcaat tagtcagcaa ccatagtccc gcccctaact
     2521 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     2581 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     2641 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     2701 attttcggag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     2761 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     2821 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2881 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     2941 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3001 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3061 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3121 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3181 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3241 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3301 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3361 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3421 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3481 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3541 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3601 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3661 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3721 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3781 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3841 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3901 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3961 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4021 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4081 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4141 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4201 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4261 taccgcgaga tccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4321 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4381 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4441 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4501 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4561 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4621 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4681 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4741 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4801 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4861 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4921 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4981 tactcat
//
