LOCUS       sequence-331742-        6170 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  mMoClo TUPV4, with CAG promoter and an MCS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6170)
  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 6170)
  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: pPD564 CAG MCS (TUPV4)
FEATURES             Location/Qualifiers
     source          1..6170
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          128..1144
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1068..1090
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1152..1171
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     1245..1264
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1245..1263
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1404..1421)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1410..1634
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2403..2831
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2490..2509)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2700..2721
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2854..3050
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2901..3036
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2963..2982
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3170..3291
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3207..3226)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3261..3280
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3340..3356)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3340..3356)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3353..3375)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3364..3380
                     /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(3388..3394,3395..3412,3413..3418))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3433..3454
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3571..3588)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3742..4330)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3822..3841)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4501..5292,5293..5361))
                     /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     5124..5143
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5362..5466)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5696..6075
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg
       61 aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg
      121 gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg
      181 ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc
      241 ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct
      301 gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg
      361 tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg
      421 tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc
      481 ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg
      541 gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc
      601 cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac
      661 ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg
      721 cggggcgggg ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag
      781 cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga
      841 gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg
      901 ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg
      961 cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg
     1021 ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg
     1081 gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct
     1141 acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattctt
     1201 atctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat
     1261 agggagtccc aagctggcta gcgtttaaac ttaagcttgg taccgagctc ggatccacta
     1321 gtccagtgtg gtggaattct gcagatatcc agcacagtgg cggccgctcg agtctagagg
     1381 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     1441 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     1501 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     1561 ggtggggcag gacagcaagg gggaggattg ggaagtcaat agcaggcatg ctggggatgc
     1621 ggtgggctct atggcttctg aggcaggtcg acactgttta acaagcccgt cagtagagcg
     1681 catcgcgata agctctagag ggacagcccc cccccaaagc ccccagggat gtaattacgt
     1741 ccctcccccg ctagggggca gcagcgagcc gcccggggct ccgctccggt ccggcgctcc
     1801 ccccgcatcc ccgagccggc agcgtgcggg gacagcccgg gcacggggaa ggtggcacgg
     1861 gatcgctttc ctctgaacgc ttctcgctgc tctttgagcc tgcagacacc tggggggata
     1921 cggggaaaaa gctttaggct gaaagagaga tttagaatga cagaatcata gaacggcctg
     1981 cgtacggatg gggctctagc tctagagctt gatatcgaat tcctgcagcc ccgctagagg
     2041 gacagccccc ccccaaagcc cccagggatg taattacgtc cctcccccgc tagggggcag
     2101 cagcgagccg cccggggctc cgctccggtc cggcgctccc cccgcatccc cgagccggca
     2161 gcgtgcgggg acagcccggg cacggggaag gtggcacggg atcgctttcc tctgaacgct
     2221 tctcgctgct ctttgagcct gcagacacct ggggggatac ggggaaaaag ctttaggctg
     2281 aaagagagat ttagaatgac agaatcatag aacggcctgg gcgcgcctgg tggcgaacga
     2341 tcaacagacg ggagaggcag aaagtcttcg ctgacagcga ccggactcta gggggtatcc
     2401 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2461 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2521 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2581 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2641 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2701 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2761 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2821 taacgcgaat taattctgtg gaatgtgtgt cagttaggct caattagtca gcaaccatag
     2881 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2941 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3001 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3061 gagcttgtat atccattttc ggaggttggg cttcggaatc gttttccggg acgccggctg
     3121 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     3181 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3241 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3301 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3361 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3421 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3481 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3541 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3601 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3661 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3721 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3781 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3841 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3901 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3961 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4021 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4081 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4141 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4201 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4261 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4321 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4381 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4441 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4501 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4561 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4621 cagtgctgca atgataccgc gagatccacg ctcaccggct ccagatttat cagcaataaa
     4681 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4741 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4801 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4861 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4921 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4981 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5041 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5101 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5161 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5221 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5281 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5341 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5401 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5461 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcgaagacg
     5521 gttacaatat cccgtgaagc ttgagtggaa tcaagatctg gggaggtcgc tgagtagtgc
     5581 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     5641 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
     5701 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     5761 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     5821 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     5881 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     5941 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     6001 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     6061 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc
     6121 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
//
