LOCUS       sequence-318353-        5161 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mMoClo TUPV, with ZF43x6-S promoter and mKate2 in the MCS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5161)
  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 5161)
  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: pHIE048 ZF43x6-S mKate2 (TUPV1)
FEATURES             Location/Qualifiers
     source          1..5161
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      455..464
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             461..1159
                     /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(1204..1221)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1210..1434
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2203..2631
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2290..2309)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2500..2521
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2654..2850
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2701..2836
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2763..2782
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2970..3091
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3007..3026)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3061..3080
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3140..3156)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3140..3156)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3153..3175)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3164..3180
                     /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(3188..3194,3195..3212,3213..3218))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3233..3254
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3371..3388)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3542..4130)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3622..3641)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4301..5092,5093..5161))
                     /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     4924..4943
                     /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 caagatcgcg tcgacggatc gggagatctg aattcggatc cagagtgagg
      241 actacctata gaagtaagag tgaggacaat cgtatacaag tagatccaga gtgaggacta
      301 cctatagaag taagagtgag gacaatcgta tacaagtaga tccagagtga ggactaccta
      361 tagaagtaag agtgaggaca atcgtataca agtagatccc tctagagggt atataatggg
      421 ggccactagt ctactaccag agctcatcgc tagcgccacc atggtgagcg agctgattaa
      481 ggagaacatg cacatgaagc tgtatatgga gggcaccgtg aacaaccacc acttcaagtg
      541 tacctccgag ggcgaaggca agccctacga gggcacccag accatgagaa tcaaggcggt
      601 cgagggcggc cctctcccct tcgccttcga catcctggct accagcttca tgtacggcag
      661 caaaaccttc atcaaccaca cccagggcat ccccgacttc tttaagcagt ccttccccga
      721 gggcttcaca tgggagagag tcaccacata cgaagatggg ggcgtgctga ccgctaccca
      781 ggacaccagc ctccaggacg gctgcctcat ctacaacgtc aagatcagag gggtgaactt
      841 cccatccaac ggccctgtga tgcagaagaa aacactcggc tgggaggcct ccaccgaaac
      901 cctgtacccc gctgacggcg gcctggaagg cagagccgac atggccctga agctcgtggg
      961 cgggggccac ctgatctgca acttgaaaac cacatacaga tccaagaaac ccgctaagaa
     1021 cctcaagatg cccggcgtct actatgtgga caggagactg gaaagaatca aggaggccga
     1081 caaagaaacc tacgtcgagc agcacgaggt ggctgtggcc agatactgcg acctccctag
     1141 caaactgggg cacagatgag cggccgctcg agtctagagg gcccgtttaa acccgctgat
     1201 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     1261 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     1321 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     1381 gggaggattg ggaagtcaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     1441 aggcaggtcg agggtacatg cgccttactc cttgtgagcg catcgcgata agctctagag
     1501 ggacagcccc cccccaaagc ccccagggat gtaattacgt ccctcccccg ctagggggca
     1561 gcagcgagcc gcccggggct ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc
     1621 agcgtgcggg gacagcccgg gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc
     1681 ttctcgctgc tctttgagcc tgcagacacc tggggggata cggggaaaaa gctttaggct
     1741 gaaagagaga tttagaatga cagaatcata gaacggcctg cgtacggatg gggctctagc
     1801 tctagagctt gatatcgaat tcctgcagcc ccgctagagg gacagccccc ccccaaagcc
     1861 cccagggatg taattacgtc cctcccccgc tagggggcag cagcgagccg cccggggctc
     1921 cgctccggtc cggcgctccc cccgcatccc cgagccggca gcgtgcgggg acagcccggg
     1981 cacggggaag gtggcacggg atcgctttcc tctgaacgct tctcgctgct ctttgagcct
     2041 gcagacacct ggggggatac ggggaaaaag ctttaggctg aaagagagat ttagaatgac
     2101 agaatcatag aacggcctgg gcgcgcctgg tggcgaacga tcaacagacg ggagagggca
     2161 aaagtcttcg ctgacagcga ccggactcta gggggtatcc ccacgcgccc tgtagcggcg
     2221 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2281 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2341 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2401 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2461 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2521 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     2581 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     2641 gaatgtgtgt cagttaggct caattagtca gcaaccatag tcccgcccct aactccgccc
     2701 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2761 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     2821 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     2881 ggaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     2941 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     3001 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3061 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     3121 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3181 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     3241 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3301 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3361 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3421 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3481 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3541 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3601 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3661 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3721 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3781 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3841 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3901 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3961 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4021 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4081 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4141 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4201 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4261 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4321 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4381 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4441 gagatccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4501 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4561 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4621 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4681 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4741 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4801 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4861 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4921 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4981 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5041 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5101 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5161 t
//
