LOCUS       sequence-318443-        6851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mMoClo TUPV, with CAG promoter and mKate2 in the MCS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6851)
  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 6851)
  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: pHIE041 CAG mKate2 (TUPV 1)
FEATURES             Location/Qualifiers
     source          1..6851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        199..578
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          854..1870
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1794..1816
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1878..1897
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     1971..1990
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1971..1989
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2009..2018
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2015..2713
                     /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(2758..2775)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2764..2988
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3757..4185
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3844..3863)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4054..4075
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4208..4404
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4255..4390
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4317..4336
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    4524..4645
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4561..4580)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4615..4634
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4694..4710)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4694..4710)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4707..4729)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4718..4734
                     /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(4742..4748,4749..4766,4767..4772))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4787..4808
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4925..4942)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5096..5684)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5176..5195)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5855..6646,6647..6715))
                     /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     6478..6497
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6716..6820)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgaag acggtgccaa ggtcttgaca aacgagtgct agtacaagat
       61 ctggggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
      121 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
      181 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      241 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      301 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      361 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      421 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      481 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      541 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct
      601 gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt
      661 aattattttg tgcagcgatg ggggcggggg gggggggggg gcgccaggcg gggcggggcg
      721 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
      781 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
      841 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc
      901 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
      961 cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct
     1021 gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc
     1081 ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg
     1141 cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg
     1201 gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg
     1261 cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca
     1321 accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct
     1381 ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt
     1441 gccgggcggg gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc
     1501 ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc
     1561 gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag
     1621 gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga
     1681 aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc
     1741 tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg
     1801 cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt
     1861 tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga
     1921 attcttatct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
     1981 cactataggg agtcccaagc tggctagcgc caccatggtg agcgagctga ttaaggagaa
     2041 catgcacatg aagctgtata tggagggcac cgtgaacaac caccacttca agtgtacctc
     2101 cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg cggtcgaggg
     2161 cggccctctc cccttcgcct tcgacatcct ggctaccagc ttcatgtacg gcagcaaaac
     2221 cttcatcaac cacacccagg gcatccccga cttctttaag cagtccttcc ccgagggctt
     2281 cacatgggag agagtcacca catacgaaga tgggggcgtg ctgaccgcta cccaggacac
     2341 cagcctccag gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcccatc
     2401 caacggccct gtgatgcaga agaaaacact cggctgggag gcctccaccg aaaccctgta
     2461 ccccgctgac ggcggcctgg aaggcagagc cgacatggcc ctgaagctcg tgggcggggg
     2521 ccacctgatc tgcaacttga aaaccacata cagatccaag aaacccgcta agaacctcaa
     2581 gatgcccggc gtctactatg tggacaggag actggaaaga atcaaggagg ccgacaaaga
     2641 aacctacgtc gagcagcacg aggtggctgt ggccagatac tgcgacctcc ctagcaaact
     2701 ggggcacaga tgagcggccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct
     2761 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2821 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2881 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2941 attgggaagt caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcag
     3001 gtcgagggta catgcgcctt actccttgtg agcgcatcgc gataagctct agagggacag
     3061 ccccccccca aagcccccag ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg
     3121 agccgcccgg ggctccgctc cggtccggcg ctccccccgc atccccgagc cggcagcgtg
     3181 cggggacagc ccgggcacgg ggaaggtggc acgggatcgc tttcctctga acgcttctcg
     3241 ctgctctttg agcctgcaga cacctggggg gatacgggga aaaagcttta ggctgaaaga
     3301 gagatttaga atgacagaat catagaacgg cctgcgtacg gatggggctc tagctctaga
     3361 gcttgatatc gaattcctgc agccccgcta gagggacagc ccccccccaa agcccccagg
     3421 gatgtaatta cgtccctccc ccgctagggg gcagcagcga gccgcccggg gctccgctcc
     3481 ggtccggcgc tccccccgca tccccgagcc ggcagcgtgc ggggacagcc cgggcacggg
     3541 gaaggtggca cgggatcgct ttcctctgaa cgcttctcgc tgctctttga gcctgcagac
     3601 acctgggggg atacggggaa aaagctttag gctgaaagag agatttagaa tgacagaatc
     3661 atagaacggc ctgggcgcgc ctggtggcga acgatcaaca gacgggagag ggcaaaagtc
     3721 ttcgctgaca gcgaccggac tctagggggt atccccacgc gccctgtagc ggcgcattaa
     3781 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3841 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3901 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3961 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4021 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4081 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     4141 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     4201 gtgtcagtta ggctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4261 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4321 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4381 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggaggt
     4441 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4501 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4561 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4621 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     4681 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4741 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4801 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4861 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4921 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4981 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5041 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5101 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5161 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5221 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5281 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5341 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5401 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5461 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5521 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5581 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5641 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5701 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5761 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5821 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5881 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5941 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagatc
     6001 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6061 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6121 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6181 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6241 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6301 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6361 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6421 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6481 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6541 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6601 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6661 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6721 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6781 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6841 cacctgacgt c
//
