LOCUS       sequence-437289-        5239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for expression PpASCL in mammalian cells  (HEK293NT).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5239)
  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 5239)
  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: pNK7186
FEATURES             Location/Qualifiers
     source          1..5239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        282..485
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     436..456
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      514..523
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1944..2065
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1981..2000)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2035..2054
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2072..2527)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2209..2230)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2421..2440
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2554..2658
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2660..3017
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2660..2851
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2683..2703)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2868..3003
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3052..3846
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3052..3846
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3106..3125)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3716..3735
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4034..4053)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4078..4125
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4454..5042
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4943..4962
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(5217..5239,1..281)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
       61 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      121 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      181 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      241 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      301 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      361 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      421 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      481 gagctggttt agtgaaccgt cagatccgct agcgccacca tggctagtcg tcgtgtcgag
      541 gcagcattcg atggacaagc agtagaactg ggtgctacta ttcctgccgc caatggcaac
      601 ggaactcatc agtcaattaa ggttcctgga catcgtcaag tgactccagg caagactacg
      661 attatggcta ttggccgtgc tgtgcccgct aacacgacgt tcaatgatgg tttggcagac
      721 cattacatcc aggagttcaa tctgcaagat ccagtccttc aggctaaact gcgtcgtttg
      781 tgcgaaacca caaccgtgaa aacgcgttat ctggtcgtca ataaggagat tctggatgag
      841 catcctgagt tcctggttga tggagctgca accgtatctc agcgtctggc tattacaggc
      901 gaagctgtta cccaacttgg acatgaggca gcaacggctg ccatcaaaga gtggggccgt
      961 ccagcctccg aaattaccca tcttgtttat gtttcctcat ccgaaatccg tctgccaggt
     1021 ggcgacttgt acctggcaca attgcttggc ttgcgttcag acgtgaaccg tgtaatgttg
     1081 tacatgctgg gttgctatgg tggtgcatct ggaatccgtg ttgccaaaga cttggctgag
     1141 aataacccag gctcacgtgt acttctgatc acgtcagaat gcactttgat tggatataag
     1201 tcactttccc ctgaccgtcc ttacgacctg gttggagcag ctctgttcgg tgacggtgct
     1261 gccgctatga ttatgggaaa agatccaatc cctgttttgg agcgtgcttt ctttgagctt
     1321 gactgggccg gtcagtcttt tattcctgga actaacaaaa ccatcgatgg tcgtttgtct
     1381 gaagaaggca tttccttcaa gctgggtcgt gaactgccta agttgattga atccaatatt
     1441 cagggtttct gcgatcctat cttgaaacgt gctggtggtc tgaagtataa cgatattttc
     1501 tgggcagttc acccaggcgg accagctatc ttgaatgctg tccagaaaca actggatttg
     1561 gcacccgaga aacttcagac cgcacgtcag gttctgcgtg actacggtaa catttctagt
     1621 tccacctgca tttacgtact ggactatatg cgtcatcaat ccttgaaact gaaggaagca
     1681 aatgacaacg taaatacgga accagagtgg ggtctgctgc ttgcatttgg ccccggcgtc
     1741 actatcgagg gtgctttgct gcgtaacctg tgctgaaagc ttcgaattct gcagtcgacg
     1801 gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc cataccacat
     1861 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     1921 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     1981 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     2041 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     2101 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     2161 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     2221 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     2281 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     2341 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     2401 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     2461 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     2521 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2581 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2641 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     2701 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2761 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2821 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     2881 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2941 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3001 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     3061 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3121 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3181 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     3241 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3301 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3361 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3421 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3481 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3541 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     3601 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3661 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3721 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3781 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     3841 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     3901 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     3961 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     4021 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     4081 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     4141 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     4201 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     4261 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     4321 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4381 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4441 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4501 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4561 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4621 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4681 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4741 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4801 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4861 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4921 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4981 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5041 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5101 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
     5161 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     5221 acataactta cggtaaatg
//
