LOCUS       sequence-325170-        5335 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRE3GV promoter and DsRedExpress2 in TUPV2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5335)
  TITLE     Leonard Lab mMoClo Plasmids
REFERENCE   2  (bases 1 to 5335)
  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"
COMMENT     Sequence Label: pPD1019 TRE3GV DsRedExpress2 in TU2
FEATURES             Location/Qualifiers
     source          1..5335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        82..431
                     /label=TRE3GV promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, optimized for 
                     retroviral and lentiviral vectors"
     protein_bind    90..108
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    126..144
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    162..180
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    198..216
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    234..252
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    270..288
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    306..324
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     370..394
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     476..495
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        476..494
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      514..523
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             520..1197
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(700..720)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1112..1135
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1242..1259)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1248..1472
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2241..2669
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2328..2347)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2538..2559
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2692..2888
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2739..2874
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2801..2820
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3008..3129
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3045..3064)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3099..3118
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3178..3194)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3178..3194)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3191..3213)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3202..3218
                     /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(3226..3232,3233..3250,3251..3256))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3271..3292
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3409..3426)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3580..4168)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3660..3679)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4339..5130,5131..5199))
                     /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     4962..4981
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5200..5304)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgaag acgggcaaaa ccgatgttga cggactaatc ctgacaagat
       61 caggcccttt catcttcact cgagtttact ccctatcagt gatagagaac gtatgaagag
      121 tttactccct atcagtgata gagaacgtat gcagacttta ctccctatca gtgatagaga
      181 acgtataagg agtttactcc ctatcagtga tagagaacgt atgaccagtt tactccctat
      241 cagtgataga gaacgtatct acagtttact ccctatcagt gatagagaac gtatatccag
      301 tttactccct atcagtgata gagaacgtat gtcgaggtag gcgtgtacgg tgggcgccta
      361 taaaagcaga gctcgtttag tgaaccgtca gatcgcctgg agcaattcca caacactttt
      421 gtcttatact tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata
      481 cgactcacta tagggagtcc caagctggct agcgccacca tggatagcac tgagaacgtc
      541 atcaagccct tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
      601 gagatcgagg gcgagggcga gggcaagccc tacgagggca cccagaccgc caagctccag
      661 gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttccagtac
      721 ggctccaagg tgtacgtgaa gcaccccgcc gacatccccg actacaagaa gctgtccttc
      781 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
      841 acccaggact cctccctcca ggacggcacc ttcatctacc acgtgaagtt catcggcgtg
      901 aacttcccct ccgacggccc cgtaatgcag aagaaaactc tgggctggga gccctccacc
      961 gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccacaaggc gctgaagctg
     1021 aagggcggcg gccactacct ggtggagttc aagtcaatct acatggccaa gaagcccgtg
     1081 aagctgcccg gctactacta cgtggactcc aagctggaca tcacctccca caacgaggac
     1141 tacaccgtgg tggagcagta cgagcgcgcc gaggcccgcc accacctgtt ccagtaggcg
     1201 gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     1261 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     1321 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     1381 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagtcaatag
     1441 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcaggtcgaa gaactatttc
     1501 ctggctgtta cgcgagcgca tcgcgataag ctctagaggg acagcccccc cccaaagccc
     1561 ccagggatgt aattacgtcc ctcccccgct agggggcagc agcgagccgc ccggggctcc
     1621 gctccggtcc ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga cagcccgggc
     1681 acggggaagg tggcacggga tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg
     1741 cagacacctg gggggatacg gggaaaaagc tttaggctga aagagagatt tagaatgaca
     1801 gaatcataga acggcctgcg tacggatggg gctctagctc tagagcttga tatcgaattc
     1861 ctgcagcccc gctagaggga cagccccccc ccaaagcccc cagggatgta attacgtccc
     1921 tcccccgcta gggggcagca gcgagccgcc cggggctccg ctccggtccg gcgctccccc
     1981 cgcatccccg agccggcagc gtgcggggac agcccgggca cggggaaggt ggcacgggat
     2041 cgctttcctc tgaacgcttc tcgctgctct ttgagcctgc agacacctgg ggggatacgg
     2101 ggaaaaagct ttaggctgaa agagagattt agaatgacag aatcatagaa cggcctgggc
     2161 gcgcctggtg gcgaacgatc aacagacggg agaggactaa agtcttcgct gacagcgacc
     2221 ggactctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2281 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2341 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2401 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2461 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2521 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2581 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2641 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttaggctca
     2701 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2761 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2821 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2881 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg aggttgggct tcggaatcgt
     2941 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     3001 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3061 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3121 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     3181 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3241 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3301 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3361 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3421 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3481 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3541 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3601 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3661 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3721 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3781 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3841 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3901 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3961 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4021 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4081 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4141 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4201 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4261 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4321 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4381 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4441 gggcttacca tctggcccca gtgctgcaat gataccgcga gatccacgct caccggctcc
     4501 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4561 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4621 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4681 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4741 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4801 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4861 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4921 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4981 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5041 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5101 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5161 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5221 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5281 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
