LOCUS       sequence-317944-        5073 bp    DNA     circular SYN 15-APR-2025
DEFINITION  mMoClo TUPV, with ZF43x3-S promoter and mKate2 in the MCS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5073)
  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 5073)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pHIE047 ZF43x3-S mKate2 (TUPV1)
FEATURES             Location/Qualifiers
     source          1..5073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      262..271
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             268..966
                     /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(1011..1028)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1017..1241
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2010..2438
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2097..2116)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2307..2328
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2461..2657
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2508..2643
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2570..2589
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2777..2898
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2814..2833)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2868..2887
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2947..2963)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2947..2963)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2960..2982)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2971..2987
                     /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(2995..3001,3002..3019,3020..3025))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3040..3061
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3178..3195)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3349..3937)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3429..3448)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4108..4899,4900..4968))
                     /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     4731..4750
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4969..5073)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcgaa gacggtgcca
       61 aggtcttgac aaacgagtgc tagtacaaga tcgcgtcgac ggatcgggag atctgaatta
      121 gagtgaggac aatcgtatac aagtagatcc agagtgagga ctacctatag aagtaagagt
      181 gaggacaatc gtatacaagt agatccctct agagggtata taatgggggc cactagtcta
      241 ctaccagagc tcatcgctag cgccaccatg gtgagcgagc tgattaagga gaacatgcac
      301 atgaagctgt atatggaggg caccgtgaac aaccaccact tcaagtgtac ctccgagggc
      361 gaaggcaagc cctacgaggg cacccagacc atgagaatca aggcggtcga gggcggccct
      421 ctccccttcg ccttcgacat cctggctacc agcttcatgt acggcagcaa aaccttcatc
      481 aaccacaccc agggcatccc cgacttcttt aagcagtcct tccccgaggg cttcacatgg
      541 gagagagtca ccacatacga agatgggggc gtgctgaccg ctacccagga caccagcctc
      601 caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttccc atccaacggc
      661 cctgtgatgc agaagaaaac actcggctgg gaggcctcca ccgaaaccct gtaccccgct
      721 gacggcggcc tggaaggcag agccgacatg gccctgaagc tcgtgggcgg gggccacctg
      781 atctgcaact tgaaaaccac atacagatcc aagaaacccg ctaagaacct caagatgccc
      841 ggcgtctact atgtggacag gagactggaa agaatcaagg aggccgacaa agaaacctac
      901 gtcgagcagc acgaggtggc tgtggccaga tactgcgacc tccctagcaa actggggcac
      961 agatgagcgg ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     1021 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     1081 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     1141 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     1201 agtcaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg caggtcgagg
     1261 gtacatgcgc cttactcctt gtgagcgcat cgcgataagc tctagaggga cagccccccc
     1321 ccaaagcccc cagggatgta attacgtccc tcccccgcta gggggcagca gcgagccgcc
     1381 cggggctccg ctccggtccg gcgctccccc cgcatccccg agccggcagc gtgcggggac
     1441 agcccgggca cggggaaggt ggcacgggat cgctttcctc tgaacgcttc tcgctgctct
     1501 ttgagcctgc agacacctgg ggggatacgg ggaaaaagct ttaggctgaa agagagattt
     1561 agaatgacag aatcatagaa cggcctgcgt acggatgggg ctctagctct agagcttgat
     1621 atcgaattcc tgcagccccg ctagagggac agcccccccc caaagccccc agggatgtaa
     1681 ttacgtccct cccccgctag ggggcagcag cgagccgccc ggggctccgc tccggtccgg
     1741 cgctcccccc gcatccccga gccggcagcg tgcggggaca gcccgggcac ggggaaggtg
     1801 gcacgggatc gctttcctct gaacgcttct cgctgctctt tgagcctgca gacacctggg
     1861 gggatacggg gaaaaagctt taggctgaaa gagagattta gaatgacaga atcatagaac
     1921 ggcctgggcg cgcctggtgg cgaacgatca acagacggga gagggcaaaa gtcttcgctg
     1981 acagcgaccg gactctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     2041 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2101 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2161 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2221 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2281 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2341 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2401 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     2461 ttaggctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     2521 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2581 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     2641 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga ggttgggctt
     2701 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     2761 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     2821 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2881 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     2941 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     3001 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     3061 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     3121 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3181 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3241 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3301 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3361 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3421 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3481 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3541 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3601 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3661 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3721 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3781 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3841 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3901 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3961 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4021 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4081 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4141 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4201 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag atccacgctc
     4261 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4321 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4381 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4441 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4501 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4561 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4621 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4681 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4741 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4801 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4861 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4921 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4981 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5041 tatttagaaa aataaacaaa taggggttcc gcg
//
