LOCUS       sequence-193240-        5476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Bnip3_Exon2+3 splice variant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5476)
  TITLE     Gordon Lab plasmids
REFERENCE   2  (bases 1 to 5476)
  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"
COMMENT     Sequence Label: HA-mus-Bnip3_Exon(2+3)
FEATURES             Location/Qualifiers
     source          1..5476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        25..404
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        405..608
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     559..579
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     653..672
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        653..671
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             697..723
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(819..837)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(820..837)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(857..874)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    863..1087
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1133..1561
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1220..1239)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1430..1451
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1570..1590)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1575..1904
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1755..1890
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1817..1836
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1971..2765
                     /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(2025..2044)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2635..2654
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2939..3060
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2976..2995)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3030..3049
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3109..3125)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3109..3125)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3122..3144)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3133..3149
                     /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(3157..3163,3164..3181,3182..3187))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3202..3223
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3340..3357)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3511..4099)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3591..3610)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4270..5061,5062..5130))
                     /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     4893..4912
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5131..5235)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5310..5329)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa
       61 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      121 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      181 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      241 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      301 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      361 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      421 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      481 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      541 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      601 gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga
      661 ctcactatag ggagacccaa gcttggtacc atggcgtacc catacgatgt tccagattac
      721 gccggatcca tcgatgtcga cgaattcatg tcgcagagcg gggagaagaa gagaagttga
      781 aagtatcctg actcgagcat gcatctagag ggccctattc tatagtgtca cctaaatgct
      841 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
      901 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
      961 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     1021 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     1081 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     1141 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1201 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     1261 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     1321 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     1381 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     1441 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     1501 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     1561 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     1621 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     1681 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     1741 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     1801 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     1861 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     1921 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     1981 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     2041 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     2101 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     2161 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     2221 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     2281 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     2341 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     2401 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     2461 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     2521 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     2581 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     2641 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     2701 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     2761 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     2821 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     2881 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     2941 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3001 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3061 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     3121 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3181 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3241 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3301 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3361 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3421 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3481 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3541 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3601 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3661 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3721 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3781 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3841 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3901 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3961 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4021 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4081 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4141 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4201 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4261 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4321 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4381 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4441 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4501 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4561 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4621 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4681 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4741 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4801 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4861 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4921 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4981 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5041 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5101 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5161 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5221 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     5281 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5341 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     5401 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
     5461 tttgcgctgc ttcgcg
//
