LOCUS       sequence-238576-        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"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1892..1948
                     /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    2097..2218
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2134..2153)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2188..2207
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2225..2680)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2362..2383)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2574..2593
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2707..2811
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2813..3170
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2813..3004
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2836..2856)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3021..3156
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3205..3999
                     /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             3205..3999
                     /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(3259..3278)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3869..3888
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4187..4206)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4231..4278
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4607..5195
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5096..5115
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5370..5673
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcatggagga
      241 cgctaagaac atcaagaagg gcccagctcc attttatcct ctggaagacg gaacagcagg
      301 agaacagctg cataaggcta tgaagagata cgctctggtg ccaggagcta ttgcttttac
      361 agacgctcac attcaggtgg acgtgacata cgccgaatac ttcgagatga gcgtgagact
      421 ggcagaagcc atgagaagat acggcctgaa taccaaccac agaattgtgg tgtgcagcga
      481 gaacagctct cagttcttca tgccagtgct gggagctctg tttattggag tggccgtggc
      541 tccagctaac gatatctata acgagagaga gctgctgaac tctatgggaa tctctcagcc
      601 tacagtggtg tttgtgagca agaagggact gagaaaagtg ctgaacgtgc agaagaagct
      661 gcccatcatc cggaagatca tcatcatgga cagcaagacc gattaccagg gattccagag
      721 catgtacacc ttcgtgacaa gccatctgcc tccttctttc aacgagtacg acttcgtgcc
      781 agagtctttc gacagagaca agacaatcgc cctgatcatg aactctagcg gatctacagg
      841 actgcctaaa ggagtggctc tgcctcatag aacagcttgc gtgagattca gccacgctag
      901 agatcctatc ttcggatacc agaacatccc agatacagcc attctgagcg tggtgccttt
      961 tcaccacgga tttggcatgt tcacaacact gggctacctg atttgcggat tcagagtggt
     1021 gctgatgtac aggttcgagg aagaactgtt tctgagaagc ctccaggact acaagattca
     1081 gtcagctctg ctggtgccta cactgttttc ttgtctggcc aagagcacac tgatcgataa
     1141 gtacgacctg agcagcctga gagaaattgc ttcaggagga gctcctctgt ctaaggaagt
     1201 gggagaagca gtggctaaga gattcagact gccaggaatc agacagggat acggactgac
     1261 agaaaccaca aacgcagtga tgatcacccc agaaggagat agaaagccag gaagcgtggg
     1321 aaaagtggtg cctttctttg aggctaaggt ggtggatctg gtgacaggaa aaaccctggg
     1381 agtgaatcag agaggagaac tgtgcgtgag aggacctatg atcatgtcag gatacgtgaa
     1441 caatccagag gccacaaacg ctctcatcga taaagacggt tggctgcata gcggagatat
     1501 cgcttattgg gacgaggacg aacacttctt catcgtggac agactgaaga gcctgatcaa
     1561 gtacaagggc tatcaggtgg ctccagcaga actggaaggc atcctgctcc agcaccctta
     1621 tatttttgac gccggagtgg caggactgcc agacgacgac gcaggagaac tgccagcagc
     1681 agtcgtggtg ctggaacacg gaaaaaccat gaccgagaag gagatcgtgg actacgtggc
     1741 ttctcaggtg acaacagcta agaaactgag aggaggagtg gtgtttgtgg acgaagtgcc
     1801 tagaggatct acaggaaagc tggacgctag aaagatcaga gagatcctga ccaaggccaa
     1861 gaaggacgga aaaattgccg tgggaagcgg agctactaac ttcagcctgc tgaagcaggc
     1921 tggagacgtg gaggagaacc ctggacctat aacccggggt acctgataac aattggcggc
     1981 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     2041 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     2101 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2161 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     2221 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     2281 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     2341 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     2401 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     2461 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     2521 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     2581 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     2641 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     2701 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2761 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     2821 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2881 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2941 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3001 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3061 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3121 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     3181 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3241 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3301 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3361 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     3421 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3481 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3541 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3601 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3661 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3721 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3781 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3841 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3901 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3961 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4021 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4081 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4141 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     4201 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     4261 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     4321 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     4381 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     4441 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4501 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4561 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4621 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4681 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4741 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4801 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4861 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4921 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4981 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5041 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5101 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5161 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5221 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5281 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     5341 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     5401 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     5461 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     5521 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     5581 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     5641 tacatctacg tattagtcat cgctattacc atg
//
