LOCUS       sequence-438205-        5310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for expression At4CL2 in mammalian cells  (HEK293NT).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5310)
  AUTHORS   Palkina KA, Karataeva TA, Perfilov MM, Fakhranurova LI, Markina NM, 
            Somermeyer LG, Garcia-Perez E, Vazquez-Vilar M, Rodriguez-Rodriguez 
            M, Vazquez-Vilriales V, Shakhova ES, Mitiouchkina T, Belozerova OA, 
            Kovalchuk SI, Alekberova A, Malyshevskaia AK, Bugaeva EN, Guglya EB,
            Balakireva A, Sytov N, Bezlikhotnova A, Boldyreva DI, Babenko VV, 
            Kondrashov FA, Choob VV, Orzaez D, Yampolsky IV, Mishin AS, 
            Sarkisyan KS
  TITLE     A hybrid pathway for self-sustained luminescence.
  JOURNAL   Sci Adv. 2024 Mar 8;10(10):eadk1992. doi: 10.1126/sciadv.adk1992. 
            Epub 2024 Mar 8.
  PUBMED    38457503
REFERENCE   2  (bases 1 to 5310)
  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: "Sci Adv."; 
            date: "2024-03-8"; volume: "10(10):eadk1992. doi"; pages: " 
            10.1126/sciadv.adk1992. Epub 2024 Mar 8"
COMMENT     Sequence Label: pNK7189
FEATURES             Location/Qualifiers
     source          1..5310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        91..1269
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          322..1260
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1217..1237
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        1553..1565
                     /gene="Pichia pastoris ADE2"
                     /label=PpADE2 HC promoter
                     /note="weak 13-bp promoter fragment that requires high-copy
                     integration for efficient expression"
     polyA_signal    2968..3079
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    3092..3112
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3113..3170
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(3194..3211)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3365..3953)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3445..3464)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4186..5001)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4186..5001)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4909..4928
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4993..5012
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     5206..5224
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(5228..5271)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    5274..5295
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ggagaagctt tgcaaagatg gataaagttt taaacagaga ggaatctttg cagctaatgg
       61 accttctagg tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc
      121 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac cggtgcctag
      181 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
      241 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
      301 gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt
      361 acgggttatg gcccttgcgt gccttgaatt acttccacct ggctgcagta cgtgattctt
      421 gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc
      481 ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc
      541 tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat ttaaaatttt
      601 tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc gggccaagat
      661 ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg tgcgtcccag
      721 cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg acgggggtag
      781 tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc
      841 tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg gccgcttccc
      901 ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg ggcgggtgag
      961 tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg tgactccacg
     1021 gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag tacgtcgtct
     1081 ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg ggtggagact
     1141 gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct ttttgagttt
     1201 ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct tccatttcag
     1261 gtgtcgtgaa atgaccacgc aagatgtaat agtaaatgat caaaatgatc aaaagcaatg
     1321 ttccaatgac gttatcttcc gatcacgttt acccgatata tatattccca atcatttgcc
     1381 cctgcacgat tacatttttg agaatatatc cgagtttgcc gccaaacctt gcctcatcaa
     1441 cgggccgaca ggagaagtct atacgtacgc agacgtccat gtcacaagcc gaaaactggc
     1501 tgctgggctc cataacttag gcgttaagca acacgacgta gtcatgattt tgcttccaaa
     1561 ctctccggaa gttgtactta ctttcttggc cgcttccttc atcggcgcca ttacaacctc
     1621 cgccaatccc ttcttcactc cggcagaaat ttccaaacag gctaaggctt ccgcagctaa
     1681 attgatcgta acacagtcac gatatgttga caaaattaag aacctccaga atgatggcgt
     1741 cttgatagta actactgatt ctgacgcaat tccagagaac tgtttgaggt tttcagagtt
     1801 gacacaaagt gaagaacccc gtgtcgactc tattccagag aaaatttccc ccgaggatgt
     1861 cgttgctctg ccgttttcat ctggtaccac agggctgcca aagggtgtaa tgctcacgca
     1921 caaaggactt gtgacttctg tcgctcaaca ggtcgatggg gaaaatccaa acctctactt
     1981 caaccgtgat gatgtgattc tctgcgtctt gccaatgttc cacatttacg ccctcaactc
     2041 aataatgctt tgctccttga gagtgggagc cacaatactc attatgccta agtttgagat
     2101 tacacttctc ctcgaacaga tacaacgatg taaggtaact gtcgctatgg tggtgccacc
     2161 gatagtgttg gccatagcaa agtccccgga gacagagaaa tatgacttat cttcagtgag
     2221 aatggtaaag tctggagcag cacctcttgg taaagaatta gaggatgcca tatccgccaa
     2281 atttcctaat gccaagttag gtcaggggta cggcatgacg gaagctggtc cggttttggc
     2341 aatgtctctc ggctttgcta aggagccttt tcccgtgaag tccggcgcat gtggaacggt
     2401 tgtgcgaaat gcagagatga agattctcga tccggacaca ggcgactcat taccgcgtaa
     2461 caagccagga gagatatgta taagagggaa ccagatcatg aagggttatc tgaatgatcc
     2521 gctggctact gcttctacta tagacaaaga tggttggctc catacggggg atgtcggctt
     2581 tatcgatgat gacgatgaat tatttatcgt cgataggctc aaagagctca tcaagtacaa
     2641 aggatttcaa gtcgcacctg ccgaattgga atccttgttg ataggtcacc ctgaaataaa
     2701 cgacgtagct gtggtagcca tgaaagaaga agatgctggg gaggttcccg ttgcctttgt
     2761 tgtcaggtcc aaggacagca acattagtga ggacgaaatc aaacaatttg tatccaaaca
     2821 ggtcgtattc tataaacgta tcaataaagt attctttacg gactctattc caaaagcacc
     2881 gagcggcaag atattaagaa aagacctgcg agccaggttg gcaaacgggt tgatgaactc
     2941 aggtagttaa ctagcttagt taactagctg tgccttctag ttgccagcca tctgttgttt
     3001 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     3061 aaaatgagga aattgcatcc gctatgtctt ctactagtag cggccgctgc agtccggcaa
     3121 aaaagggcaa ggtgtcacca ccctgccctt tttctttaaa accgaaaaga ttacttcgcg
     3181 ttatgcaggc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3241 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3301 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3361 cgtttttcca caggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3421 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3481 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3541 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3601 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3661 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3721 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3781 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3841 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3901 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3961 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4021 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4081 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagctcgagt cccgtcaagt
     4141 cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga aaaactcatc
     4201 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     4261 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     4321 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     4381 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     4441 tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     4501 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     4561 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     4621 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     4681 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     4741 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     4801 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     4861 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     4921 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctgg agcaagacgt
     4981 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     5041 tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     5101 aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcagc tcgagtgcca
     5161 cttgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     5221 aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt ctggaattcg
     5281 cggccgcttc tagagactag tggaagacat
//
