LOCUS       sequence-193239-        5660 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Bnip3_deltaExon3 splice variant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5660)
  AUTHORS   Diehl-Jones W, Archibald A, Gordon JW, Mughal W, Hossain Z, Friel JK
  TITLE     Human Milk Fortification Increases Bnip3 Expression Associated With 
            Intestinal Cell Death In Vitro.
  JOURNAL   J Pediatr Gastroenterol Nutr. 2015 Nov;61(5):583-90. doi: 
            10.1097/MPG.0000000000000876.
  PUBMED    26505960
REFERENCE   2  (bases 1 to 5660)
  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: "10.1097/MPG"; 
            journalName: "J Pediatr Gastroenterol Nutr"; date: "2015-11"; 
            volume: "61"; issue: "5"; pages: "583-90"
COMMENT     Sequence Label: HA-mus-Bnip3_deltaExon3
FEATURES             Location/Qualifiers
     source          1..5660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..406
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        407..610
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     561..581
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     655..674
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        655..673
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             699..725
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1005..1023)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1006..1023)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1043..1060)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1049..1273
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1319..1747
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1406..1425)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1616..1637
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1756..1776)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1761..2090
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1941..2076
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2003..2022
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2157..2951
                     /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(2211..2230)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2821..2840
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3125..3246
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3162..3181)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3216..3235
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3295..3311)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3295..3311)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3308..3330)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3319..3335
                     /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(3343..3349,3350..3367,3368..3373))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3388..3409
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3526..3543)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3697..4285)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3777..3796)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4456..5247,5248..5316))
                     /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     5079..5098
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5317..5421)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5496..5515)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      421 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      481 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      541 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      601 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
      661 gactcactat agggagaccc aagcttggta ccatggcgta cccatacgat gttccagatt
      721 acgccggatc catcgatgtc gacgaattca tgtcgcagag cggggaggag aacctgcagg
      781 gctcctgggt agaactgcac ttcagcaatg gcaatgggag cagcgttcca gcctccgtct
      841 ctatttataa tggtgacatg gaaaaaatac tgttggatgc ccagcatgaa tctggacgaa
      901 gtagctccaa gagttctcac tgtgacagtc tgaggaagat tatattgaga gaagaagaga
      961 agttgaaagt atcctgactc gagcatgcat ctagagggcc ctattctata gtgtcaccta
     1021 aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     1081 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1141 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     1201 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     1261 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     1321 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1381 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1441 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1501 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1561 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1621 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1681 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1741 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     1801 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     1861 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     1921 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     1981 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     2041 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     2101 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     2161 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     2221 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     2281 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     2341 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     2401 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     2461 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     2521 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     2581 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     2641 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     2701 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     2761 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     2821 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     2881 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     2941 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     3001 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3061 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3121 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3181 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3241 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     3301 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3361 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3421 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3481 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3541 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3601 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3661 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3721 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3781 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3841 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3901 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3961 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4021 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4081 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4141 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4201 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4261 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4321 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4381 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4441 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4501 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4561 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4621 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4681 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4741 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4801 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4861 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4921 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4981 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5041 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5101 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5161 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5221 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5281 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5341 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5401 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     5461 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     5521 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     5581 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
     5641 aggcgttttg cgctgcttcg
//
