LOCUS       sequence-356523-        6273 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EOLA1/CXORF40A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6273)
  TITLE     ASCH domain proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6273)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pcDNA3.1-EOLA1/CXORF40A
FEATURES             Location/Qualifiers
     source          1..6273
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1376..1486
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="DEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     CDS             1493..1516
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(1826..1844)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1827..1844)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1864..1881)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1870..2094
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2140..2568
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2227..2246)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2437..2458
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2577..2597)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2582..2911
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2762..2897
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2824..2843
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2978..3772
                     /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(3032..3051)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3642..3661
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3946..4067
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3983..4002)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4037..4056
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4116..4132)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4116..4132)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4129..4151)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4140..4156
                     /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(4164..4170,4171..4188,4189..4194))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4209..4230
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4347..4364)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4518..5106)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4598..4617)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5277..6068,6069..6137))
                     /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     5900..5919
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6138..6242)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccag ccatgaagtt
      901 cggctgtctg agctttagac agccctatgc cggattcgtg ctgaacggca tcaagaccgt
      961 ggagaccaga tggagacctc tgctgagcag ccagaggaac tgcaccatcg ccgtgcacat
     1021 cgcccataga gactgggagg gagacgcttg gagggagctg ctggtggaga gactgggcat
     1081 gacccccgcc cagattcaag ctctgctgag gaagggcgag aagttcggaa gaggcgtgat
     1141 cgccggactg gtggatatcg gcgagacact gcagtgcccc gaggatctga cccccgacga
     1201 ggtggtggag ctggagaacc aagccgtgct gaccaatctg aagcagaagt acctcaccgt
     1261 gatcagcaac cccagatggc tgctggaacc cattcctaga aagggcggca aggacgtgtt
     1321 ccaagtggat atccccgagc atctgattcc tctgggccac gaggtgagca gcggcgacga
     1381 gaagaccacc ggctggagag gcggccacgt ggtcgagggg ctagccggtg agctggagca
     1441 gctacgggcc cgccttgagc accaccccca ggggcagcgc gagccgggat ccgactacaa
     1501 agacgatgac gataagtgac cggtgaactt cccgccgccg ttgttgtttt ggagcacgga
     1561 aagacgatga cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa
     1621 aagttgcgcg gaggagttgt gtttgtggac gaagtaccga aaggtcttac cggaaaactc
     1681 gacgcaagaa aaatcagaga gatcctcata aaggccaaga agggcggaaa gtccaaattg
     1741 taaaatgtaa ctgtattcag cgatgacgaa attcttagct attgtaatcc tccgaggcct
     1801 cgagcatgca tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat
     1861 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     1921 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     1981 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2041 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     2101 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     2161 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2221 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2281 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2341 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2401 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2461 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     2521 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     2581 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2641 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2701 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2761 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2821 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     2881 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     2941 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3001 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3061 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3121 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     3181 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3241 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3301 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3361 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3421 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3481 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     3541 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3601 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     3661 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     3721 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     3781 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     3841 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     3901 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     3961 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4021 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4081 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4141 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4201 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4261 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4321 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4381 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4441 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4501 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4561 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4621 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4681 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4741 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4801 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4861 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4921 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4981 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5041 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5101 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5161 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5221 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5281 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5341 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5401 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5461 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5521 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5581 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5641 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5701 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5761 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5821 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5881 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5941 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6001 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6061 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6121 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6181 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6241 cgcacatttc cccgaaaagt gccacctgac gtc
//
