LOCUS       sequence-317523-        8584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP-BIRC3 with the short 3'UTR in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8584)
  AUTHORS   Lee SH, Mayr C
  TITLE     Gain of Additional BIRC3 Protein Functions through 3'-UTR-Mediated 
            Protein Complex Formation.
  JOURNAL   Mol Cell. 2019 May 16;74(4):701-712.e9. doi: 
            10.1016/j.molcel.2019.03.006. Epub 2019 Apr 1.
  PUBMED    30948266
REFERENCE   2  (bases 1 to 8584)
  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: 
            "10.1016/j.molcel.2019.03.006"; journalName: "Mol Cell"; date: 
            "2019-05-16- 16"; volume: "74"; issue: "4"; pages: "701-712.e9"
COMMENT     Sequence Label: pcDNA-GFP-BIRC3-SU
FEATURES             Location/Qualifiers
     source          1..8584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(701..703,704..706,707..1417)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(746..767)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1007..1026)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1354..1375
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3941..3958)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3947..4171
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4217..4645
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4304..4323)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4514..4535
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4654..4674)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4659..4988
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4839..4974
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4901..4920
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5055..5849
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5109..5128)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5719..5738
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6023..6144
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6060..6079)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6114..6133
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6193..6209)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6193..6209)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6206..6228)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6217..6233
                     /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(6241..6247,6248..6265,6266..6271))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6286..6307
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6424..6441)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6595..7183)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6675..6694)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7354..8145,8146..8214))
                     /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     7977..7996
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8215..8319)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8394..8413)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc atggtgagca agggcgagga
      721 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      781 gttcagcgtg tctggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
      841 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
      901 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
      961 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1021 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1081 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1141 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg cgaacttcaa
     1201 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     1261 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     1321 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1381 cgccgggatc actctcggca tggacgagct gtacaagaac atagtagaaa acagcatatt
     1441 cttatcaaat ttgatgaaaa gcgccaacac gtttgaactg aaatacgact tgtcatgtga
     1501 actgtaccga atgtctacgt attccacttt tcctgctggg gttcctgtct cagaaaggag
     1561 tcttgctcgt gctggtttct attacactgg tgtgaatgac aaggtcaaat gcttctgttg
     1621 tggcctgatg ctggataact ggaaaagagg agacagtcct actgaaaagc ataaaaagtt
     1681 gtatcctagc tgcagattcg ttcagagtct aaattccgtt aacaacttgg aagctacctc
     1741 tcagcctact tttccttctt cagtaacaaa ttccacacac tcattacttc cgggtacaga
     1801 aaacagtgga tatttccgtg gctcttattc aaactctcca tcaaatcctg taaactccag
     1861 agcaaatcaa gatttttctg ccttgatgag aagttcctac cactgtgcaa tgaataacga
     1921 aaatgccaga ttacttactt ttcagacatg gccattgact tttctgtcgc caacagatct
     1981 ggcaaaagca ggcttttact acataggacc tggagacaga gtggcttgct ttgcctgtgg
     2041 tggaaaattg agcaattggg aaccgaagga taatgctatg tcagaacacc tgagacattt
     2101 tcccaaatgc ccatttatag aaaatcagct tcaagacact tcaagataca cagtttctaa
     2161 tctgagcatg cagacacatg cagcccgctt taaaacattc tttaactggc cctctagtgt
     2221 tctagttaat cctgagcagc ttgcaagtgc gggtttttat tatgtgggta acagtgatga
     2281 tgtcaaatgc ttttgctgtg atggtggact caggtgttgg gaatctggag atgatccatg
     2341 ggttcaacat gccaagtggt ttccaaggtg tgagtacttg ataagaatta aaggacagga
     2401 gttcatccgt caagttcaag ccagttaccc tcatctactt gaacagctgc tatccacatc
     2461 agacagccca ggagatgaaa atgcagagtc atcaattatc cattttgaac ctggagaaga
     2521 ccattcagaa gatgcaatca tgatgaatac tcctgtgatt aatgctgccg tggaaatggg
     2581 ctttagtaga agcctggtaa aacagacagt tcagagaaaa atcctagcaa ctggagagaa
     2641 ttatagacta gtcaatgatc ttgtgttaga cttactcaat gcagaagatg aaataaggga
     2701 agaggagaga gaaagagcaa ctgaggaaaa agaatcaaat gatttattat taatccggaa
     2761 gaatagaatg gcactttttc aacatttgac ttgtgtaatt ccaatcctgg atagtctact
     2821 aactgccgga attattaatg aacaagaaca tgatgttatt aaacagaaga cacagacgtc
     2881 tttacaagca agagaactga ttgatacgat tttagtaaaa ggaaatattg cagccactgt
     2941 attcagaaac tctctgcaag aagctgaagc tgtgttatat gagcatttat ttgtgcaaca
     3001 ggacataaaa tatattccca cagaagatgt ttcagatcta ccagtggaag aacaattgcg
     3061 gagactacaa gaagaaagaa catgtaaagt gtgtatggac aaagaagtgt ccatagtgtt
     3121 tattccttgt ggtcatctag tagtatgcaa agattgtgct ccttctttaa gaaagtgtcc
     3181 tatttgtagg agtacaatca agggtacagt tcgtacattt ctttcatgag agctcagaag
     3241 aaccaaaaca tcgtctaaac tttagaatta atttattaaa tgtattataa ctttaacttt
     3301 tatcctaatt tggtttcctt aaaattttta tttatttaca actcaaaaaa cattgttttg
     3361 tgtaacatat ttatatatgt atctaaacca tatgaacata tattttttag aaactaagag
     3421 aatgataggc ttttgttctt atgaacgaaa aagaggtagc actacaaaca caatattcaa
     3481 tcaaaatttc agcattattg aaattgtaag tgaagtaaaa cttaagatat ttgagttaac
     3541 ctttaagaat tttaaatatt ttggcattgt actaataccg ggaacatgaa gccaggtgtg
     3601 gtggtatgtg cctgtagtcc caggctgagg caagagaatt acttgagccc aggagtttga
     3661 atccatcctg ggcagcatac tgagaccctg cctttaaaaa caaacagaac aaaaacaaaa
     3721 caccagggac acatttctct gtcttttttg atcagtgtcc tatacatcga aggtgtgcat
     3781 atatgttgaa tgacatttta gggacatggt gtttttataa agaattctgt gagaaaaaat
     3841 ttaataaagc aacaaaaatt actcttattc ttcattgctt taactagtgt tctagagcgg
     3901 ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     3961 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     4021 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     4081 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     4141 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     4201 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4261 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4321 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4381 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4441 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4501 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4561 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4621 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4681 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4741 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4801 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4861 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4921 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4981 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     5041 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5101 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     5161 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     5221 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5281 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5341 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5401 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5461 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5521 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5581 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5641 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5701 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5761 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5821 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5881 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5941 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     6001 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     6061 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6121 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     6181 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     6241 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     6301 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6361 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6421 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6481 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6541 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6601 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6661 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6721 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6781 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6841 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6901 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6961 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7021 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7081 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7141 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     7201 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     7261 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     7321 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     7381 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     7441 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     7501 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7561 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7621 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7681 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7741 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7801 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7861 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7921 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7981 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     8041 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     8101 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     8161 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     8221 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     8281 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     8341 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
     8401 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     8461 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     8521 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
     8581 cgtt
//
