LOCUS       sequence-317926-        5324 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mMoClo TUPV, with ZF43x12-S promoter and mKate2 in the MCS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5324)
  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 5324)
  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: pHIE049 ZF43x12-S mKate2 (TUPV1)
FEATURES             Location/Qualifiers
     source          1..5324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      505..514
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             511..1209
                     /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(1254..1271)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1260..1484
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2253..2681
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2340..2359)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2550..2571
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2704..2900
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2751..2886
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2813..2832
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3020..3141
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3057..3076)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3111..3130
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3190..3206)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3190..3206)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3203..3225)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3214..3230
                     /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(3238..3244,3245..3262,3263..3268))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3283..3304
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3421..3438)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3592..4180)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3672..3691)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4351..5142,5143..5211))
                     /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     4974..4993
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5212..5316)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgac gtcgacggat cgggagatcg aagacggtgc caaggtcttg
       61 acaaacgagt gctagtacaa gatctgaatt cggatccaga gtgaggacta cctatagaag
      121 taagagtgag gacaatcgta tacaagtaga tccagagtga ggactaccta tagaagtaag
      181 agtgaggaca atcgtataca agtagatcca gagtgaggac tacctataga agtaagagtg
      241 aggacaatcg tatacaagta gatccctcta gagggagatc cagagtgagg actacctata
      301 gaagtaagag tgaggacaat cgtatacaag tagatccaga gtgaggacta cctatagaag
      361 taagagtgag gacaatcgta tacaagtaga tccagagtga ggactaccta tagaagtaag
      421 agtgaggaca atcgtataca agtagatccc tctagagggt atataatggg ggccactagt
      481 ctactaccag agctcatcgc tagcgccacc atggtgagcg agctgattaa ggagaacatg
      541 cacatgaagc tgtatatgga gggcaccgtg aacaaccacc acttcaagtg tacctccgag
      601 ggcgaaggca agccctacga gggcacccag accatgagaa tcaaggcggt cgagggcggc
      661 cctctcccct tcgccttcga catcctggct accagcttca tgtacggcag caaaaccttc
      721 atcaaccaca cccagggcat ccccgacttc tttaagcagt ccttccccga gggcttcaca
      781 tgggagagag tcaccacata cgaagatggg ggcgtgctga ccgctaccca ggacaccagc
      841 ctccaggacg gctgcctcat ctacaacgtc aagatcagag gggtgaactt cccatccaac
      901 ggccctgtga tgcagaagaa aacactcggc tgggaggcct ccaccgaaac cctgtacccc
      961 gctgacggcg gcctggaagg cagagccgac atggccctga agctcgtggg cgggggccac
     1021 ctgatctgca acttgaaaac cacatacaga tccaagaaac ccgctaagaa cctcaagatg
     1081 cccggcgtct actatgtgga caggagactg gaaagaatca aggaggccga caaagaaacc
     1141 tacgtcgagc agcacgaggt ggctgtggcc agatactgcg acctccctag caaactgggg
     1201 cacagatgag cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac
     1261 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     1321 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     1381 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     1441 ggaagtcaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcaggtcg
     1501 agggtacatg cgccttactc cttgtgagcg catcgcgata agctctagag ggacagcccc
     1561 cccccaaagc ccccagggat gtaattacgt ccctcccccg ctagggggca gcagcgagcc
     1621 gcccggggct ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc agcgtgcggg
     1681 gacagcccgg gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc ttctcgctgc
     1741 tctttgagcc tgcagacacc tggggggata cggggaaaaa gctttaggct gaaagagaga
     1801 tttagaatga cagaatcata gaacggcctg cgtacggatg gggctctagc tctagagctt
     1861 gatatcgaat tcctgcagcc ccgctagagg gacagccccc ccccaaagcc cccagggatg
     1921 taattacgtc cctcccccgc tagggggcag cagcgagccg cccggggctc cgctccggtc
     1981 cggcgctccc cccgcatccc cgagccggca gcgtgcgggg acagcccggg cacggggaag
     2041 gtggcacggg atcgctttcc tctgaacgct tctcgctgct ctttgagcct gcagacacct
     2101 ggggggatac ggggaaaaag ctttaggctg aaagagagat ttagaatgac agaatcatag
     2161 aacggcctgg gcgcgcctgg tggcgaacga tcaacagacg ggagagggca aaagtcttcg
     2221 ctgacagcga ccggactcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2281 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2341 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2401 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2461 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2521 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2581 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2641 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     2701 cagttaggct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2761 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2821 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2881 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggaggttggg
     2941 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     3001 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     3061 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3121 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     3181 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3241 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3301 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3361 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3421 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3481 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3541 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3601 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3661 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3721 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3781 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3841 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3901 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3961 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4021 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4081 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4141 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4201 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4261 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4321 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4381 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4441 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagatccacg
     4501 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4561 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4621 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4681 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4741 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4801 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4861 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4921 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4981 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5041 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5101 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5161 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5221 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5281 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttc
//
