LOCUS       sequence-317949-        5140 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mMoClo TUPV, with ZF43x12-C promoter and mKate2 in the MCS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5140)
  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 5140)
  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: pHIE046 ZF43x12-C mKate2 (TUPV1)
FEATURES             Location/Qualifiers
     source          1..5140
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      467..895
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(554..573)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     764..785
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        918..1114
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      965..1100
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1027..1046
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    1234..1355
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1271..1290)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1325..1344
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1404..1420)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1404..1420)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1417..1439)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1428..1444
                     /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(1452..1458,1459..1476,1477..1482))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1497..1518
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1635..1652)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1806..2394)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1886..1905)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2565..3356,3357..3425))
                     /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     3188..3207
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3426..3530)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      3859..3868
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3865..4563
                     /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(4608..4625)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4614..4838
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 ggctgaaaga gagatttaga atgacagaat catagaacgg cctgcgtacg gatggggctc
       61 tagctctaga gcttgatatc gaattcctgc agccccgcta gagggacagc ccccccccaa
      121 agcccccagg gatgtaatta cgtccctccc ccgctagggg gcagcagcga gccgcccggg
      181 gctccgctcc ggtccggcgc tccccccgca tccccgagcc ggcagcgtgc ggggacagcc
      241 cgggcacggg gaaggtggca cgggatcgct ttcctctgaa cgcttctcgc tgctctttga
      301 gcctgcagac acctgggggg atacggggaa aaagctttag gctgaaagag agatttagaa
      361 tgacagaatc atagaacggc ctgggcgcgc ctggtggcga acgatcaaca gacgggagag
      421 ggcaaaagtc ttcgctgaca gcgaccggac tctagggggt atccccacgc gccctgtagc
      481 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
      541 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
      601 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
      661 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
      721 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
      781 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
      841 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
      901 tgtggaatgt gtgtcagtta ggctcaatta gtcagcaacc atagtcccgc ccctaactcc
      961 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     1021 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     1081 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     1141 tttcggaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     1201 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     1261 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1321 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     1381 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1441 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     1501 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1561 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1621 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1681 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1741 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1801 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1861 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1921 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1981 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2041 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2101 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2161 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2221 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2281 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2341 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2401 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2461 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2521 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2581 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2641 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2701 ccgcgagatc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2761 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     2821 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     2881 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     2941 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3001 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3061 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3121 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3181 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3241 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3301 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3361 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3421 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3481 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3541 cgaaaagtgc cacctgacgt cgacggatcg ggagatcgaa gacggtgcca aggtcttgac
     3601 aaacgagtgc tagtacaaga tcggagtgag gaaaattaga gtgaggacga tcggagtgag
     3661 gataattaga gtgaggacga tcggagtgag gaaaattaga gtgaggacga tcggagtgag
     3721 gataattaga gtgaggacga tcggagtgag gaaaattaga gtgaggacga tcggagtgag
     3781 gataattaga gtgaggacga tccctctaga gggtatataa tgggggccac tagtctacta
     3841 ccagagctca tcgctagcgc caccatggtg agcgagctga ttaaggagaa catgcacatg
     3901 aagctgtata tggagggcac cgtgaacaac caccacttca agtgtacctc cgagggcgaa
     3961 ggcaagccct acgagggcac ccagaccatg agaatcaagg cggtcgaggg cggccctctc
     4021 cccttcgcct tcgacatcct ggctaccagc ttcatgtacg gcagcaaaac cttcatcaac
     4081 cacacccagg gcatccccga cttctttaag cagtccttcc ccgagggctt cacatgggag
     4141 agagtcacca catacgaaga tgggggcgtg ctgaccgcta cccaggacac cagcctccag
     4201 gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcccatc caacggccct
     4261 gtgatgcaga agaaaacact cggctgggag gcctccaccg aaaccctgta ccccgctgac
     4321 ggcggcctgg aaggcagagc cgacatggcc ctgaagctcg tgggcggggg ccacctgatc
     4381 tgcaacttga aaaccacata cagatccaag aaacccgcta agaacctcaa gatgcccggc
     4441 gtctactatg tggacaggag actggaaaga atcaaggagg ccgacaaaga aacctacgtc
     4501 gagcagcacg aggtggctgt ggccagatac tgcgacctcc ctagcaaact ggggcacaga
     4561 tgagcggccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     4621 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     4681 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     4741 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaagt
     4801 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcag gtcgagggta
     4861 catgcgcctt actccttgtg agcgcatcgc gataagctct agagggacag ccccccccca
     4921 aagcccccag ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg
     4981 ggctccgctc cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc
     5041 ccgggcacgg ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg
     5101 agcctgcaga cacctggggg gatacgggga aaaagcttta
//
