LOCUS       sequence-218025-        4859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expressing tandem cDNAs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4859)
  TITLE     Michael E Wright Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4859)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pTRE-T2-∆mIR-IRES-Puromycin
FEATURES             Location/Qualifiers
     source          1..4859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        952..1266
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    960..978
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    996..1014
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1031..1049
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1067..1085
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1103..1121
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1138..1156
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1174..1192
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1239..1263
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    1309..1881
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1476..1493)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     polyA_signal    2021..2102
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2205..2227
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2265..2283)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2351..2455
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2456..2524,2525..3316)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGARGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2674..2693)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(3457..3477)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3462..3791
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3642..3777
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3884..4483
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3884..3903)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4380..4400
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4713..4794
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4804..4823
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggct ggagtttact
      961 ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgatg
     1021 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
     1081 agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttatcc
     1141 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
     1201 cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt gaaccgtcag
     1261 atcgcctgga gaattgatcg aattcgatcg cggccgcgat cacgcgtgcc cctctccctc
     1321 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     1381 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     1441 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     1501 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     1561 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     1621 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     1681 gatagttgtg gaaagagtca aatggctcac ctcaagcgta ttcaacaagg ggctgaagga
     1741 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
     1801 atgtgtttag tcgaggttaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     1861 ctttgaaaaa cacgatgata atggtaccga tcactagtga tctctagact agaggatcat
     1921 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     1981 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2041 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2101 gccactagag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     2161 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     2221 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     2281 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     2341 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2401 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2461 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2521 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2581 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2641 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     2701 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2761 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2821 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2881 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2941 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3001 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3061 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3121 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg gggctcgcgg
     3181 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3241 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3301 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3361 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3421 aatcccttaa cgtgagtttt cgttccagct agctgtggaa tgtgtgtcag ttagggtgtg
     3481 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3541 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3601 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3661 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3721 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3781 gcttttgcaa aaagcttgca tgcctgcagg tcggccgcca cgaccggtgc cgccaccatc
     3841 ccctgaccca cgcccctgac ccctcacaag gagacgacct tccatgaccg agtacaagcc
     3901 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggct gtacgcaccc tcgccgccgc
     3961 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt
     4021 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     4081 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     4141 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     4201 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     4261 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     4321 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggaaacct ccgcgccccg
     4381 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     4441 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc
     4501 gcagcgcccg accgaaagga gcgcacgacc ccatggctcc gaccgaagcc gacccgggcg
     4561 gccccgccga ccccgcaccc gcccccgagg cccaccgact caagaggatc ataatcagcc
     4621 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     4681 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     4741 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4801 gttgtggttt gtccaaactc atcaatgtat ctggccgtag aaaagatcaa aggatcttc
//
