LOCUS       sequence-306894-        6884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dox/Tet-inducible expression of dominant negative CREB (A-CREB) in 
            mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6884)
  AUTHORS   Musicant AM, Parag-Sharma K, Gong W, Sengupta M, Chatterjee A, Henry
            EC, Tsai YH, Hayward MC, Sheth S, Betancourt R, Hackman TG, Padilla 
            RJ, Parker JS, Giudice J, Flaveny CA, Hayes DN, Amelio AL
  TITLE     CRTC1/MAML2 directs a PGC-1alpha-IGF-1 circuit that confers 
            vulnerability to PPARgamma inhibition.
  JOURNAL   Cell Rep. 2021 Feb 23;34(8):108768. doi: 
            10.1016/j.celrep.2021.108768.
  PUBMED    33626346
REFERENCE   2  (bases 1 to 6884)
  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: "Cell Rep.";
            date: "2021-02-23"; volume: "34(8):108768. doi"; pages: " 
            10.1016/j.celrep.2021.108768"
COMMENT     Sequence Label: pRetroX-Tight_A-CREB_PGK-tdTomato
FEATURES             Location/Qualifiers
     source          1..6884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(80..579)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(128..147)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     521..544
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     CDS             complement(829..861)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             complement(865..888)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(919..1233)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     primer_bind     complement(922..946)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    993..1011
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1029..1047
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1064..1082
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1100..1118
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1136..1154
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1171..1189
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1207..1225
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             complement(1269..1685)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(1285..1301)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(1317..1339)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    1886..2085
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             2148..2323
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     promoter        complement(2324..2527)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        2528..2907
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3007..3025)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3093..3197
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3198..3266,3267..4058)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3416..3435)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4229..4817
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4718..4737
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4971..4988
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5069..5088)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        complement(5110..5439)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5124..5259
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5424..5444
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     5580..5602
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     LTR             5705..6130
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     CDS             complement(join(6248..6884,1..65))
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(6331..6354)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(6529..6548)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6798..6816
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
ORIGIN
        1 gagccctcca tgcgcacctt gaagcgcatg aactctttga tgacctcctc gcccttgctc
       61 accatcttgg gctgcaggtc gaaaggcccg gagatgagga agaggagaac agcgcggcag
      121 acgtgcgctt ttgaagcgtg cagaatgccg ggcctccgga ggaccttcgg gcgcccgccc
      181 cgcccctgag cccgcccctg agcccgcccc cggacccacc ccttcccagc ctctgagccc
      241 agaaagcgaa ggagcaaagc tgctattggc cgctgcccca aaggcctacc cgcttccatt
      301 gctcagcggt gctgtccatc tgcacgagac tagtgagacg tgctacttcc atttgtcacg
      361 tcctgcacga cgcgagctgc ggggcggggg ggaacttcct gactagggga ggagtagaag
      421 gtggcgcgaa ggggccacca aagaacggag ccggttggcg cctaccggtg gatgtggaat
      481 gtgtgcgagg ccagaggcca cttgtgtagc gccaagtgcc cagcggggct gctaaagcgc
      541 atgctccaga ctgccttggg aaaagcgcct cccctacccg gtagaattca cgcgttcgcg
      601 atctagagcc ggcgcggccg cctctagaag cttgaattaa tctgacttgt ggcagtaaag
      661 gtccttaagt gcttttagct cctcaatcaa tgttttgttt tggttttcaa gcactgccac
      721 tctgttctcg agttctgcca gttcctgctc cagctcttcg gcctcttttt ccagctcttc
      781 gttttcacgg gccagttcct cagcacgttg ttccaggtca ggatcccgac ccatttgctg
      841 tccaccagtc atgctagcca tatgcttgtc atcgtcgtcc ttgtagtcca tggtgggtgg
      901 cagatcgatc cttctccagg cgatctgacg gttcactaaa cgagctctgc ttatataggc
      961 ctcccaccgt acacgcctac ctcgacatac gttctctatc actgataggg agtaaactcg
     1021 acatacgttc tctatcactg atagggataa actcgacata cgttctctat cactgatagg
     1081 gagtaaactc gacatacgtt ctctatcact gatagggagt aaactcgaca tacgttctct
     1141 atcactgata gggagtaaac tcgacatcgt tctctatcac tgatagggag taaactcgac
     1201 atacgttctc tatcactgat agggagtaaa ctcgagtgaa gacgaaaggg cctcagatct
     1261 tcaattccgg cgcctagaga aggagtgagg gctggataaa gggaggatcg aggcggggtc
     1321 gaacgaggag gttcaagggg gagagacggg gcggatggag gaagaggagg cggaggctta
     1381 gggtgtacaa agggcttgac ccagggaggg gggtcaaaag ccaaggcttc ccaggtcacg
     1441 atgtagggga cctggtctgg gtgtccatgc gggccaggtg aaaagacctt gatcttaacc
     1501 tgggtgatga ggtctcggtt aaaggtgccg tctcgcggcc atccgacgtt aaaggttggc
     1561 cattctgcag agcagaaggt aacccaacgt ctcttcttga catctaccga ctggttgtga
     1621 gcgatccgct cgacatcttt ccagtgacct aaggtcaaac ttaagggagt ggtaacagtc
     1681 tggccctaat tttcagacaa atacagaaac acagtcagac agagacaaca cagaacgatg
     1741 ctgcagcgct gcagcagaca agacgcgcgg cttcggttcc aaaccgaaag caaaaattca
     1801 gacggaggcg ggaactgttt taggttctcg tctcctacca gaaccacata tcctgacggg
     1861 gtcggattcc acatcgactc ccttcctcag gtcgggccac aaaaacggcc cccaaagtcc
     1921 ctgggacgtc tcccagggtt gcggccgggt gttcagaact cgtcagttcc accacgggtc
     1981 cgccagatac agagctagtt agctaactag tacagacgca ggcgcataac atcaaacata
     2041 gacactagac aatcggacag acacagataa gttgctggcc agcttacctc ccggtggtgg
     2101 gtcggtggtc cctgggcagg ggtctccaaa tcccggacga gcccccaaat gaaagacccc
     2161 cgtcgtgggt agtcaatcac tcagaggaga ccctcccaag gaacagcgag accacgattc
     2221 ggatgcaaac agcaagaggc tttattggga atacgggtac ccgggcgacg cagtctatcg
     2281 gaagactggc gcgccgagtg aggggttgtg ggctctttta ttgagctctg cttatataga
     2341 cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt
     2401 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga
     2461 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
     2521 gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     2581 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     2641 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     2701 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     2761 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     2821 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
     2881 attaataact agtcaataat caatgtcaac atggcggtaa tgttggacat gagccaatat
     2941 aaatgtacat attatgatat ggatacaacg tatgcaatgg ccaagctaat tcttgaagac
     3001 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     3061 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3121 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3181 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3241 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3301 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3361 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3421 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     3481 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3541 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3601 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3661 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3721 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3781 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3841 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3901 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3961 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4021 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4081 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4141 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4201 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4261 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4321 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     4381 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4441 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4501 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4561 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4621 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4681 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4741 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4801 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4861 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4921 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4981 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     5041 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5101 agtaagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     5161 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     5221 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     5281 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     5341 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     5401 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagcc ggatctacac
     5461 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     5521 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5581 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagccagc
     5641 ttacctcccg gtggtgggtc ggtggtccct gggcaggggt ctcccgatcc cggacgagcc
     5701 cccaaatgaa agacccccgc tgacgggtag tcaatcactc agaggagacc ctcccaagga
     5761 acagcgagac cacaagtcgg atgcaactgc aagagggttt attggataca cgggtacccg
     5821 ggcgactcag tcaatcggag gactggcgcc ccgagtgagg ggttgtgggc tcttttattg
     5881 agctcgggga gcagaagcgc gcgaacagaa gcgagaagcg aactgattgg ttagttcaaa
     5941 taaggcacag ggtcatttca ggtccttggg gcaccctgga aacatctgat ggttctctag
     6001 ggtcgaccct gttccatctg ttcctgacct tgatctgaac ttctctattc tcagttatgt
     6061 atttttccat gccttgcaaa atggcgttac ttaagctagc ttgccaaacc tacaggtggg
     6121 gtctttcatt cccccctttt tctggagact aaataaaatc ttttatttta tctatggctc
     6181 gtactctata ggcttcagct ggtgatattg ttgagtcaaa actagagcct ggaccactga
     6241 tatctcactt gtacagctcg tccatgccgt acaggaacag gtggtggcgg ccctcggagc
     6301 gctcgtactg ttccacgatg gtgtagtcct cgttgtggga ggtgatgtcc agcttggtgt
     6361 ccacgtagta gtagccgggc agttgcacgg gcttcttggc catgtagatg gtcttgaact
     6421 ccaccaggta gtggccgccg tccttcagct tcagggcctg gtggatctcg cccttcagca
     6481 cgccgtcgcg ggggtacagg cgctcggtgg aggcctccca gcccatggtc ttcttctgca
     6541 ttacggggcc gtcggggggg aagttggtgc cgcgcatctt caccttgtag atcagcgtgc
     6601 cgtcctgcag ggaggagtcc tgggtcacgg tcaccagacc gccgtcctcg aagttcatca
     6661 cgcgctccca cttgaagccc tcggggaagg acagcttctt gtaatcgggg atgtcggcgg
     6721 ggtgcttcac gtacgccttg gagccgtaca tgaactgggg ggacaggatg tcccaggcga
     6781 agggcagggg gccgcccttg gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc
     6841 ggccctcgcc ctcgccctcg atctcgaact cgtggccgtt catg
//
