LOCUS       sequence-417796-        7215 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human codon-optimized light-driven sodium pump ErNaR 
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7215)
  AUTHORS   Podoliak E, Lamm GHU, Marin E, Schellbach AV, Fedotov DA, Stetsenko 
            A, Asido M, Maliar N, Bourenkov G, Balandin T, Baeken C, Astashkin 
            R, Schneider TR, Bateman A, Wachtveitl J, Schapiro I, Busskamp V, 
            Guskov A, Gordeliy V, Alekseev A, Kovalev K
  TITLE     A subgroup of light-driven sodium pumps with an additional Schiff 
            base counterion.
  JOURNAL   Nat Commun. 2024 Apr 10;15(1):3119. doi: 10.1038/s41467-024-47469-0.
  PUBMED    38600129
REFERENCE   2  (bases 1 to 7215)
  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: "Nat 
            Commun."; date: "2024-04-10"; volume: "15(1):3119. doi"; pages: " 
            10.1038/s41467-024-47469-0"
COMMENT     Sequence Label: pcDNA3.1(-)-C2C1-ErNaR-TS-EYFP-ES
FEATURES             Location/Qualifiers
     source          1..7215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    229..350
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(266..285)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     320..339
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(399..415)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(399..415)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(412..434)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    423..439
                     /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(447..453,454..471,472..477))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    492..513
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(630..647)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(801..1389)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(881..900)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1560..2351,2352..2420))
                     /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     2183..2202
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2421..2525)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2600..2619)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2791..3170
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3171..3374
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3325..3345
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3419..3438
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3419..3437
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3539..3548
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4595..4597,4598..5308)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4637..4658)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4898..4917)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5245..5266
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5362..5379)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5368..5592
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5638..6066
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5725..5744)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5935..5956
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6075..6095)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6080..6409
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6260..6395
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6322..6341
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             join(6476..7215,1..55)
                     /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
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDKFF"
     primer_bind     complement(6530..6549)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7140..7159
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacaagttct tctgagcggg
       61 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
      121 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
      181 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
      241 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
      301 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
      361 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
      421 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
      481 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
      541 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
      601 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
      661 cggctgcggc gagcggtatc agctcactca aaggcggtaa tatggttatc cacagaatca
      721 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      781 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      841 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      901 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      961 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1021 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1081 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1141 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1201 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     1261 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1321 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1381 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1441 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1501 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     1561 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1621 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     1681 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     1741 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     1801 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     1861 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     1921 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     1981 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     2041 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     2101 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     2161 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     2221 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     2281 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     2341 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2401 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2461 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     2521 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     2581 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     2641 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     2701 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     2761 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
     2821 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     2881 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     2941 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     3001 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     3061 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     3121 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     3181 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     3241 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     3301 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     3361 atataagcag agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta
     3421 atacgactca ctatagggag acccaagctg gctagcgttt aaacgggccc tctagactcg
     3481 agtggccgcc actgtgctgg atatctgcag aattccacca cactggacta gtggatccgc
     3541 caccatggac tatgggggag cactgtccgc cgtcgggctg tttcagactt cctacaccct
     3601 ggagaacaac gggagcgtca tttgcattcc taacaatggc cagtgcttct gtctggcctg
     3661 gctgaagagc aacggtaccc cttccatcga gaactttctg gcctacgact tctggcagta
     3721 cgacgtgatc agacacctgt tcgccttcag caccgccgtg tttctggccg gcctggtgta
     3781 ctttgccatg accgccagaa ccaccgctcc taactacaga ctgagcgcca acatcagcgc
     3841 cgtggtcatg gtgtctgctg ccctggaact gggacaactg tggctgctgt ggaacgagag
     3901 ctttcagtgg gccgagctgc agggctcttt tgtgcctgtt gccggcgaga gattcagcaa
     3961 cggctaccgg tacatgaact ggctgatcga cgtgcccatg ctggccacac agctggtggt
     4021 tgtgtgtggc tttgtgggca ccgagctgag aaacagatgg gccaagctga caatcgccgg
     4081 cgtgctgatg atcctgacag gctacgtggg ccagtacttc gaacctgctg ttgctggcgt
     4141 gccaggatat gaaggcgccg agcagttttg gatctgggga atcatctcca ccgccttttt
     4201 cgtgtggatg ctgctgattc tggccaacgc cgtgcggaat cctcaaggcg ctccttctga
     4261 tgaagtgcgg agccggctga agttctgctt ctggttcctg ctggctacct ggtctatcta
     4321 ccccttcgcc tacgccatgc ctctgtttgc cccaacagct gatggcgtgg tcgtgcggca
     4381 agtgatctat acagtggccg acgtgtccag caagctggtg tttggcgtga tcctgtctca
     4441 ggtggccctg agaagatctg ccgaggaagg ctttgagcct gccagagtgg ccagcggcga
     4501 gttcgatgag agagcaccag ctagagcggc cgccaagagc aggatcacca gcgagggcga
     4561 gtacatcccc ctggaccaga tcgacatcaa cgtggtgagc aagggcgagg agctgttcac
     4621 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     4681 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     4741 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgca
     4801 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     4861 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     4921 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     4981 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     5041 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     5101 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     5161 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa
     5221 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     5281 cactctcggc atggacgagc tgtacaagtt ctgctacgag aacgaggtgt gatgaaagct
     5341 taagtttaaa ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     5401 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     5461 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     5521 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     5581 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     5641 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5701 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     5761 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5821 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     5881 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5941 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     6001 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     6061 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     6121 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     6181 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     6241 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     6301 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     6361 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     6421 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     6481 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     6541 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     6601 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     6661 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     6721 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     6781 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     6841 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     6901 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     6961 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     7021 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     7081 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     7141 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     7201 gctttacggt atcgc
//
