LOCUS       sequence-90457-e        6057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty lentiviral expression vector for ultradestabilized Venus that 
            indicates the activity of a transcriptional element of interest.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6057)
  AUTHORS   Wang CC, Bajikar SS, Jamal L, Atkins KA, Janes KA
  TITLE     A time- and matrix-dependent TGFBR3-JUND-KRT5 regulatory circuit in 
            single breast epithelial cells and basal-like premalignancies.
  JOURNAL   Nat Cell Biol. 2014 Apr;16(4):345-56. doi: 10.1038/ncb2930. Epub 
            2014 Mar 23.
  PUBMED    24658685
REFERENCE   2  (bases 1 to 6057)
  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"; doi: "10.1038/ncb2930"; 
            journalName: "Nat Cell Biol"; date: "2014-04"; volume: "16"; issue: 
            "4"; pages: "345-56"
COMMENT     Sequence Label: pTRF.1-mCMV-udsVenus
FEATURES             Location/Qualifiers
     source          1..6057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        133..336
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     287..307
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        1410..1448
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1445..1469
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1560..1569
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1566..1568,1569..1571,1572..2282,2283..2285,2286..2288,
                     2289..2411)
                     /codon_start=1
                     /product="EYFP destabilized by residues 422-461 of mouse 
                     ornithine decarboxylase, giving an in vivo half-life of ~2 
                     hours"
                     /label=d2EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKDISHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     primer_bind     complement(1611..1632)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1872..1891)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2219..2240
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2417..3005
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2470..2490)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2888..2899)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3341..3462
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3378..3397)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3432..3451
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3481..3616
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3654..3670)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3654..3670)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3667..3689)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3678..3694
                     /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(3702..3708,3709..3726,3727..3732))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3747..3768
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3885..3902)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4056..4644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4136..4155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4815..5606,5607..5675))
                     /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     5438..5457
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5676..5780)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5848..5866
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5904..5926)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6026..6045
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
       61 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      121 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      181 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      241 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      301 aggcgtgtac ggtgggaggt ctatataagc agagcttgtg aaacttcgag gagtctcttt
      361 gttgaggact tttgagttct cccttgaggc tcccacagat acaataaata tttgagattg
      421 aaccctgtcg agtatctgtg taatcttttt tacctgtgag gtctcggaat ccgggccgag
      481 aacttcgcag ttggcgcccg aacagggact tgattgagag tgattgagga agtgaagcta
      541 gagcaataga aagctgttaa gcagaactcc tgctgaccta aatagggaag cagtagcaga
      601 cgctgctaac agtgagtatc tctagtgaag cagactcgag ctcataatca agtcattgtt
      661 taaaggccca gataaattac atctggtgac tcttcgcgga ccttcaagcc aggagattcg
      721 ccgagggaca gtcaacaagg taggagagat tctacagcaa catggggaat ggacaggggc
      781 gagattggaa aatggccatt aagagatgta gtaatgttgc tgtaggagta ggggggaaga
      841 gtaaaaaatt tggagaaggg aatttcagat gggccattag aatggctaat gtatctacag
      901 gacgagaacc tggtgatata ccagagactt tagatcaact aaggttggtt atttgcgatt
      961 tacaagaaag aagagaaaaa tttggatcta gcaaagaaat tgatatggca attcctgcat
     1021 tgaggagaaa tggtaggcaa tgtggcatgt ctgaaaaaga ggaggaatga tgaagtatct
     1081 cagacttatt ttataaggga gatactgtgc tgagttcttc cctttgagga aggtatgtca
     1141 tatcctagac atagtctcaa ttttaaaaga agaggtagga taggagggat ggccccttat
     1201 gaattattag cacaacaaga atccttaaga atacaagatt atttttctgc aataccacaa
     1261 aaattgcaag cacagtggat ttattataaa gatcaaaaag ataagaaatg gaaaggacca
     1321 atgagagtag aatactgggg acagggatca gtattattaa aggatgaaga gaagggatat
     1381 tttcttataa tcgatgaatt ctagaactag ttaggcgtgt acggtgggag gtctatataa
     1441 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     1501 tccatagaag acaccgggac cgatccagcc taagcttggt cgacaggtat ccaccggtcg
     1561 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1621 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     1681 cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc
     1741 ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt cgcccgctac cccgaccaca
     1801 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1861 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1921 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1981 ggcacaagct ggagtacaac tacaatagcc acaacgtcta tatcaccgcc gacaagcaga
     2041 agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc
     2101 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     2161 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     2221 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     2281 aggatatcag ccatggcttc ccgccggcgg tggcggcgca ggatgatggc acgctgccca
     2341 tgtcttgtgc ccaggagagc gggatggacc gtcaccctgc agcctgtgct tctgctagga
     2401 tcaatgtgta gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     2461 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2521 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     2581 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     2641 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     2701 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     2761 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     2821 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     2881 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     2941 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     3001 cccgccggta ccgatgacag agttagaaga tcgcttcagg aagctatttg gcacgacttc
     3061 tacaacggga gacagcacag tagattctga agatgaacct cctaaaaaag aaaaaagggt
     3121 ggactgggat gagtattgga accctgaaat cgatagcttc cagtgctttg tgaaacttcg
     3181 aggagtctct ttgttgagga cttttgagtt ctcccttgag gctcccacag atacaataaa
     3241 tatttgagat tgaaccctgt cgagtatctg tgtaatcttt tttacctgtg aggtctcgga
     3301 atccgggccg agaacttcgc agcgagctca ttgtaccgcg aacttgttta ttgcagctta
     3361 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3421 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     3481 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3541 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3601 ggcttttttg gaggcctaga cttttgcaga gacggcccaa attcgtaatc atggtcatag
     3661 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3721 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3781 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3841 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3901 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3961 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4021 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4081 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4141 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4201 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4261 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4321 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4381 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4441 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     4501 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4561 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4621 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4681 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4741 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4801 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4861 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4921 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4981 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5041 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5101 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5161 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5221 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5281 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5341 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5401 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5461 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5521 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5581 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5641 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5701 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5761 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     5821 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     5881 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     5941 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     6001 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcac
//
