LOCUS       sequence-228455-        5673 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Akaluc luciferase mammalian expression in -N1 backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5673)
  AUTHORS   Karlsson J, Tzeng SY, Hemmati S, Luly KM, Choi O, Rui Y, Wilson DR, 
            Kozielski KL, Quinones-Hinojosa A, Green JJ
  TITLE     Photocrosslinked Bioreducible Polymeric Nanoparticles for Enhanced 
            Systemic siRNA Delivery as Cancer Therapy.
  JOURNAL   Adv Funct Mater. 2021 Apr 22;31(17). doi: 10.1002/adfm.202009768. 
            Epub 2021 Feb 22.
  PUBMED    34650390
REFERENCE   2  (bases 1 to 5673)
  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: "Adv Funct 
            Mater."; date: "2021-04-22"; volume: "31(17). doi"; pages: " 
            10.1002/adfm.202009768. Epub 2021 Feb 22"
COMMENT     Sequence Label: Akaluc-N1
FEATURES             Location/Qualifiers
     source          1..5673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2256..2312
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     polyA_signal    2461..2582
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2498..2517)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2552..2571
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2589..3044)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2726..2747)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2938..2957
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3071..3175
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3177..3534
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3177..3368
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3200..3220)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3385..3520
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3569..4363
                     /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             3569..4363
                     /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(3623..3642)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4233..4252
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4551..4570)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4595..4642
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4971..5559
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5460..5479
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgg
      601 aggacgctaa gaacatcaag aagggcccag ctccatttta tcctctggaa gacggaacag
      661 caggagaaca gctgcataag gctatgaaga gatacgctct ggtgccagga gctattgctt
      721 ttacagacgc tcacattcag gtggacgtga catacgccga atacttcgag atgagcgtga
      781 gactggcaga agccatgaga agatacggcc tgaataccaa ccacagaatt gtggtgtgca
      841 gcgagaacag ctctcagttc ttcatgccag tgctgggagc tctgtttatt ggagtggccg
      901 tggctccagc taacgatatc tataacgaga gagagctgct gaactctatg ggaatctctc
      961 agcctacagt ggtgtttgtg agcaagaagg gactgagaaa agtgctgaac gtgcagaaga
     1021 agctgcccat catccggaag atcatcatca tggacagcaa gaccgattac cagggattcc
     1081 agagcatgta caccttcgtg acaagccatc tgcctccttc tttcaacgag tacgacttcg
     1141 tgccagagtc tttcgacaga gacaagacaa tcgccctgat catgaactct agcggatcta
     1201 caggactgcc taaaggagtg gctctgcctc atagaacagc ttgcgtgaga ttcagccacg
     1261 ctagagatcc tatcttcgga taccagaaca tcccagatac agccattctg agcgtggtgc
     1321 cttttcacca cggatttggc atgttcacaa cactgggcta cctgatttgc ggattcagag
     1381 tggtgctgat gtacaggttc gaggaagaac tgtttctgag aagcctccag gactacaaga
     1441 ttcagtcagc tctgctggtg cctacactgt tttcttgtct ggccaagagc acactgatcg
     1501 ataagtacga cctgagcagc ctgagagaaa ttgcttcagg aggagctcct ctgtctaagg
     1561 aagtgggaga agcagtggct aagagattca gactgccagg aatcagacag ggatacggac
     1621 tgacagaaac cacaaacgca gtgatgatca ccccagaagg agatagaaag ccaggaagcg
     1681 tgggaaaagt ggtgcctttc tttgaggcta aggtggtgga tctggtgaca ggaaaaaccc
     1741 tgggagtgaa tcagagagga gaactgtgcg tgagaggacc tatgatcatg tcaggatacg
     1801 tgaacaatcc agaggccaca aacgctctca tcgataaaga cggttggctg catagcggag
     1861 atatcgctta ttgggacgag gacgaacact tcttcatcgt ggacagactg aagagcctga
     1921 tcaagtacaa gggctatcag gtggctccag cagaactgga aggcatcctg ctccagcacc
     1981 cttatatttt tgacgccgga gtggcaggac tgccagacga cgacgcagga gaactgccag
     2041 cagcagtcgt ggtgctggaa cacggaaaaa ccatgaccga gaaggagatc gtggactacg
     2101 tggcttctca ggtgacaaca gctaagaaac tgagaggagg agtggtgttt gtggacgaag
     2161 tgcctagagg atctacagga aagctggacg ctagaaagat cagagagatc ctgaccaagg
     2221 ccaagaagga cggaaaaatt gccgtgggaa gcggagctac taacttcagc ctgctgaagc
     2281 aggctggaga cgtggaggag aaccctggac ctataacccg gggtacctga taacaattgg
     2341 cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     2401 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     2461 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2521 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2581 taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     2641 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     2701 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     2761 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     2821 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     2881 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     2941 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3001 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     3061 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3121 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     3181 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3241 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3301 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3361 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3421 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3481 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     3541 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3601 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3661 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3721 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     3781 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3841 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3901 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3961 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4021 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4081 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4141 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4201 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4261 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4321 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4381 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4441 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4501 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     4561 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4621 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     4681 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     4741 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     4801 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     4861 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4921 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4981 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5041 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5101 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     5161 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5221 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5281 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5341 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5401 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5461 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5521 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5581 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5641 gattctgtgg ataaccgtat taccgccatg cat
//
