LOCUS       sequence-325172-        5377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRE3GV promoter and EYFP in TUPV3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5377)
  TITLE     Leonard Lab mMoClo Plasmids
REFERENCE   2  (bases 1 to 5377)
  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: pJM467 TRE3GV EYFP in TU3
FEATURES             Location/Qualifiers
     source          1..5377
                     /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             join(520..522,523..525,526..1239)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(565..586)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(826..845)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1173..1194
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1284..1301)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1290..1514
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2283..2711
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2370..2389)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2580..2601
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2734..2930
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2781..2916
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2843..2862
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3050..3171
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3087..3106)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3141..3160
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3220..3236)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3220..3236)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3233..3255)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3244..3260
                     /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(3268..3274,3275..3292,3293..3298))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3313..3334
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3451..3468)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3622..4210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3702..3721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4381..5172,5173..5241))
                     /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     5004..5023
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5242..5346)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgaag acggactaaa tagtagttca gacgccgtta agcgcaagat
       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 tggtgagcaa gggcgaggag
      541 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      601 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
      661 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac
      721 ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
      781 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
      841 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
      901 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
      961 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     1021 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1081 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc
     1141 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1201 gccgggatca ctctcggcat ggacgagctg tacaagtagg cggccgctcg agtctagagg
     1261 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     1321 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     1381 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     1441 ggtggggcag gacagcaagg gggaggattg ggaagtcaat agcaggcatg ctggggatgc
     1501 ggtgggctct atggcttctg aggcaggtcg aagagactgc ccgacacatc ttagtgagcg
     1561 catcgcgata agctctagag ggacagcccc cccccaaagc ccccagggat gtaattacgt
     1621 ccctcccccg ctagggggca gcagcgagcc gcccggggct ccgctccggt ccggcgctcc
     1681 ccccgcatcc ccgagccggc agcgtgcggg gacagcccgg gcacggggaa ggtggcacgg
     1741 gatcgctttc ctctgaacgc ttctcgctgc tctttgagcc tgcagacacc tggggggata
     1801 cggggaaaaa gctttaggct gaaagagaga tttagaatga cagaatcata gaacggcctg
     1861 cgtacggatg gggctctagc tctagagctt gatatcgaat tcctgcagcc ccgctagagg
     1921 gacagccccc ccccaaagcc cccagggatg taattacgtc cctcccccgc tagggggcag
     1981 cagcgagccg cccggggctc cgctccggtc cggcgctccc cccgcatccc cgagccggca
     2041 gcgtgcgggg acagcccggg cacggggaag gtggcacggg atcgctttcc tctgaacgct
     2101 tctcgctgct ctttgagcct gcagacacct ggggggatac ggggaaaaag ctttaggctg
     2161 aaagagagat ttagaatgac agaatcatag aacggcctgg gcgcgcctgg tggcgaacga
     2221 tcaacagacg ggagaggtta caagtcttcg ctgacagcga ccggactcta gggggtatcc
     2281 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2341 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2401 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2461 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2521 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2581 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2641 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2701 taacgcgaat taattctgtg gaatgtgtgt cagttaggct caattagtca gcaaccatag
     2761 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2821 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     2881 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     2941 gagcttgtat atccattttc ggaggttggg cttcggaatc gttttccggg acgccggctg
     3001 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     3061 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3121 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3181 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3241 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3301 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3361 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3421 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3481 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3541 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3601 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3661 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3721 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3781 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3841 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3901 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3961 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4021 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4081 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4141 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4201 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4261 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4321 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4381 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4441 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4501 cagtgctgca atgataccgc gagatccacg ctcaccggct ccagatttat cagcaataaa
     4561 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4621 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4681 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4741 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4801 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4861 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4921 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4981 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5041 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5101 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5161 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5221 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5281 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5341 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
