LOCUS       sequence-317524-        7972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP-BIRC3 without the endogenous 3'UTR in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7972)
  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 7972)
  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-NU
FEATURES             Location/Qualifiers
     source          1..7972
                     /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(3329..3346)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3335..3559
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3605..4033
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3692..3711)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3902..3923
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4042..4062)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4047..4376
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4227..4362
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4289..4308
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4443..5237
                     /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(4497..4516)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5107..5126
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5411..5532
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5448..5467)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5502..5521
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5581..5597)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5581..5597)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5594..5616)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5605..5621
                     /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(5629..5635,5636..5653,5654..5659))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5674..5695
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5812..5829)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5983..6571)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6063..6082)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6742..7533,7534..7602))
                     /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     7365..7384
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7603..7707)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7782..7801)
                     /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 agctcggatc
     3241 cactagtcca gtgtggtgga attctgcaga tatccagcac agtggcggcc gctcgagtct
     3301 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3361 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3421 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3481 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3541 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3601 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3661 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3721 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3781 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3841 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3901 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3961 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4021 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4081 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4141 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4201 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4261 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4321 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4381 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     4441 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4501 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4561 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4621 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4681 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4741 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4801 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4861 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4921 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4981 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5041 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5101 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5161 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5221 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5281 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5341 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5401 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5461 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5521 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5581 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5641 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5701 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5761 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5821 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5881 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5941 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6001 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6061 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6121 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6181 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6241 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6301 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6361 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6421 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6481 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6541 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6601 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6661 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6721 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6781 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6841 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6901 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6961 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7021 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7081 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7141 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7201 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7261 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7321 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7381 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7441 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7501 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7561 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7621 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7681 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     7741 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     7801 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     7861 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     7921 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg tt
//
