LOCUS       sequence-324917-        5079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mMoClo TUPV4, with CMV promoter and an MCS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5079)
  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 5079)
  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: pPD474 CMV MCS (TUPV4)
FEATURES             Location/Qualifiers
     source          1..5079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        199..578
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        579..782
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     733..753
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     827..846
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        827..845
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(986..1003)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    992..1216
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1985..2413
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2072..2091)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2282..2303
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2436..2632
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2483..2618
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2545..2564
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2752..2873
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2789..2808)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2843..2862
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2922..2938)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2922..2938)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2935..2957)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2946..2962
                     /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(2970..2976,2977..2994,2995..3000))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3015..3036
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3153..3170)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3324..3912)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3404..3423)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4083..4874,4875..4943))
                     /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     4706..4725
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4944..5048)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgaag acggttacaa tatcccgtga agcttgagtg gaatcaagat
       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 gatgcggttt tggcagtaca
      601 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      661 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      721 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      781 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
      841 atagggagtc ccaagctggc tagcgtttaa acttaagctt ggtaccgagc tcggatccac
      901 tagtccagtg tggtggaatt ctgcagatat ccagcacagt ggcggccgct cgagtctaga
      961 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     1021 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     1081 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     1141 ggggtggggc aggacagcaa gggggaggat tgggaagtca atagcaggca tgctggggat
     1201 gcggtgggct ctatggcttc tgaggcaggt cgacactgtt taacaagccc gtcagtagag
     1261 cgcatcgcga taagctctag agggacagcc cccccccaaa gcccccaggg atgtaattac
     1321 gtccctcccc cgctaggggg cagcagcgag ccgcccgggg ctccgctccg gtccggcgct
     1381 ccccccgcat ccccgagccg gcagcgtgcg gggacagccc gggcacgggg aaggtggcac
     1441 gggatcgctt tcctctgaac gcttctcgct gctctttgag cctgcagaca cctgggggga
     1501 tacggggaaa aagctttagg ctgaaagaga gatttagaat gacagaatca tagaacggcc
     1561 tgcgtacgga tggggctcta gctctagagc ttgatatcga attcctgcag ccccgctaga
     1621 gggacagccc ccccccaaag cccccaggga tgtaattacg tccctccccc gctagggggc
     1681 agcagcgagc cgcccggggc tccgctccgg tccggcgctc cccccgcatc cccgagccgg
     1741 cagcgtgcgg ggacagcccg ggcacgggga aggtggcacg ggatcgcttt cctctgaacg
     1801 cttctcgctg ctctttgagc ctgcagacac ctggggggat acggggaaaa agctttaggc
     1861 tgaaagagag atttagaatg acagaatcat agaacggcct gggcgcgcct ggtggcgaac
     1921 gatcaacaga cgggagaggc agaaagtctt cgctgacagc gaccggactc tagggggtat
     1981 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2041 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2101 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2161 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2221 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2281 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2341 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2401 tttaacgcga attaattctg tggaatgtgt gtcagttagg ctcaattagt cagcaaccat
     2461 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     2521 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     2581 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     2641 gggagcttgt atatccattt tcggaggttg ggcttcggaa tcgttttccg ggacgccggc
     2701 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     2761 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2821 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     2881 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     2941 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     3001 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3061 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3121 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3181 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3241 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3301 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3361 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3421 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3481 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3541 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3601 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3661 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3721 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3781 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3841 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3901 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3961 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4021 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4081 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4141 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4201 cccagtgctg caatgatacc gcgagatcca cgctcaccgg ctccagattt atcagcaata
     4261 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4321 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4381 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4441 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4501 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4561 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4621 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4681 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4741 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4801 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4861 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4921 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4981 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5041 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
