LOCUS       sequence-325169-        4728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRE3GV promoter and an MCS in TUPV1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4728)
  TITLE     Leonard Lab mMoClo Plasmids
REFERENCE   2  (bases 1 to 4728)
  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: pPD765 TRE3GV MCS in TU1
FEATURES             Location/Qualifiers
     source          1..4728
                     /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"
     primer_bind     complement(635..652)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    641..865
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1634..2062
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1721..1740)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1931..1952
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2085..2281
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2132..2267
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2194..2213
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2401..2522
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2438..2457)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2492..2511
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2571..2587)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2571..2587)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2584..2606)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2595..2611
                     /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(2619..2625,2626..2643,2644..2649))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2664..2685
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2802..2819)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2973..3561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3053..3072)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3732..4523,4524..4592))
                     /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     4355..4374
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4593..4697)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgaag acggtgccaa ggtcttgaca aacgagtgct agtacaagat
       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 agcgtttaaa cttaagcttg gtaccgagct
      541 cggatccact agtccagtgt ggtggaattc tgcagatatc cagcacagtg gcggccgctc
      601 gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
      661 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
      721 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
      781 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagtcaa tagcaggcat
      841 gctggggatg cggtgggctc tatggcttct gaggcaggtc gagggtacat gcgccttact
      901 ccttgtgagc gcatcgcgat aagctctaga gggacagccc ccccccaaag cccccaggga
      961 tgtaattacg tccctccccc gctagggggc agcagcgagc cgcccggggc tccgctccgg
     1021 tccggcgctc cccccgcatc cccgagccgg cagcgtgcgg ggacagcccg ggcacgggga
     1081 aggtggcacg ggatcgcttt cctctgaacg cttctcgctg ctctttgagc ctgcagacac
     1141 ctggggggat acggggaaaa agctttaggc tgaaagagag atttagaatg acagaatcat
     1201 agaacggcct gcgtacggat ggggctctag ctctagagct tgatatcgaa ttcctgcagc
     1261 cccgctagag ggacagcccc cccccaaagc ccccagggat gtaattacgt ccctcccccg
     1321 ctagggggca gcagcgagcc gcccggggct ccgctccggt ccggcgctcc ccccgcatcc
     1381 ccgagccggc agcgtgcggg gacagcccgg gcacggggaa ggtggcacgg gatcgctttc
     1441 ctctgaacgc ttctcgctgc tctttgagcc tgcagacacc tggggggata cggggaaaaa
     1501 gctttaggct gaaagagaga tttagaatga cagaatcata gaacggcctg ggcgcgcctg
     1561 gtggcgaacg atcaacagac gggagagggc aaaagtcttc gctgacagcg accggactct
     1621 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     1681 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     1741 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     1801 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     1861 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     1921 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     1981 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     2041 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggc tcaattagtc
     2101 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     2161 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     2221 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     2281 aaagctcccg ggagcttgta tatccatttt cggaggttgg gcttcggaat cgttttccgg
     2341 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     2401 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2461 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2521 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     2581 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     2641 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     2701 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     2761 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2821 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2881 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2941 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3001 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3061 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3121 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3181 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3241 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3301 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3361 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3421 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3481 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3541 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3601 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3661 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3721 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3781 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3841 ccatctggcc ccagtgctgc aatgataccg cgagatccac gctcaccggc tccagattta
     3901 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3961 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4021 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4081 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4141 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4201 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4261 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4321 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4381 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4441 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4501 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4561 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4621 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4681 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
