LOCUS       sequence-459313-        6881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a rabbit IgH signal peptide fused to GFP in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6881)
  AUTHORS   Jaroenlak P, McCarty KL, Xia B, Lam C, Zwack EE, Yanai I, Bhabha G, 
            Ekiert DC
  TITLE     scRNA-seq reveals transcriptional dynamics of Encephalitozoon 
            intestinalis parasites in human macrophages.
  JOURNAL   bioRxiv [Preprint]. 2024 May 30:2024.05.30.596468. doi: 
            10.1101/2024.05.30.596468.
  PUBMED    38853846
REFERENCE   2  (bases 1 to 6881)
  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"; journalName: "bioRxiv 
            [Preprint]. 2024 May 30:2024.05.30.596468. doi: 
            10.1101/2024.05.30.596468."
COMMENT     Sequence Label: pBEL2837
FEATURES             Location/Qualifiers
     source          1..6881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        47..426
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        427..630
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     581..601
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     627..651
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             853..1566
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             853..1563
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(880..908)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1503..1523
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    1655..2243
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1708..1728)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2126..2137)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2264..2283)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2308..2356
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2558..2986
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2645..2664)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2855..2876
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2995..3015)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3000..3329
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3180..3315
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3242..3261
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3396..4190
                     /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(3450..3469)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4060..4079
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4366..4487
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4403..4422)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4457..4476
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4536..4552)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4536..4552)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4549..4571)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4560..4576
                     /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(4584..4590,4591..4608,4609..4614))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4629..4650
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4767..4784)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4938..5526)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5018..5037)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5697..6488,6489..6557))
                     /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     6320..6339
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6558..6662)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6737..6756)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
       61 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      121 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      181 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      241 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      301 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      361 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      421 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      481 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      541 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      601 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga
      661 cgccatccac gctgttttga cctccataga agacaccggg accgatccag cctccggact
      721 ctagaggatc gaacccttgg atctctagcg aattccctct agacacagac gctcaccatg
      781 gagactgggc tgcgctggct tctcctggtc gctgtgctca aaggtgtcca gtgtcagtcc
      841 ctggaggagt ccagcaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa
      901 ttagatggtg atgttaatgg gcacaaattt tctgtccgtg gagagggtga aggtgatgct
      961 acaaacggaa aactcaccct taaatttatt tgcactactg gaaaactacc tgttccgtgg
     1021 ccaacacttg tcactactct gacctatggt gttcaatgct tttcccgtta tccggatcac
     1081 atgaaacggc atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaacgcact
     1141 atatctttca aagatgacgg gacctacaag acgcgtgctg aagtcaagtt tgaaggtgat
     1201 acccttgtta atcgtatcga gttaaagggt attgatttta aagaagatgg aaacattctt
     1261 ggacacaaac tcgagtacaa ctttaactca cacaatgtat acatcacggc agacaaacaa
     1321 aagaatggaa tcaaagctaa cttcaaaatt cgccacaacg ttgaagatgg ttccgttcaa
     1381 ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac
     1441 aaccattacc tgtcgacaca atctgtcctt tcgaaagatc ccaacgaaaa gcgtgaccac
     1501 atggtccttc ttgagtttgt aactgctgct gggattacac atggcatgga tgagctctac
     1561 aaatgagcgg ccgctcgagg ccggcaaggc cggatccccc gacctcgaca agggttcgat
     1621 ccctaccggt tagtaatgag tttgatatct cgacaatcaa cctctggatt acaaaatttg
     1681 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     1741 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     1801 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     1861 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     1921 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     1981 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     2041 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     2101 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     2161 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     2221 ctccctttgg gccgcctccc cgcctggaac gggggaggct aactgaaaca cggaaggaga
     2281 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt
     2341 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     2401 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     2461 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     2521 gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2581 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2641 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2701 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2761 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2821 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2881 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2941 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3001 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3061 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3121 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3181 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3241 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3301 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3361 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3421 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3481 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3541 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3601 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3661 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3721 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3781 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3841 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3901 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     3961 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4021 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4081 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4141 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4201 ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4261 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4321 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4381 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4441 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4501 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4561 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4621 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4681 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4741 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4801 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4861 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4921 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4981 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5041 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5101 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5161 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5221 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5281 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5341 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5401 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5461 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5521 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5581 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5641 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5701 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5761 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5821 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5881 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5941 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6001 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6061 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6121 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6181 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6241 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6301 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6361 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6421 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6481 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6541 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6601 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6661 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     6721 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     6781 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     6841 aaggcttgac cgacaattgc atgaagaatc tgcttagggt t
//
