LOCUS       sequence-474609-        5573 bp    DNA     circular SYN 05-JUN-2025
DEFINITION  Express ABCA1 in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5573)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5573)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pNL [NlucP/ABCA1/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..5573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      835..844
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             841..1353
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             1357..1476
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    1528..1649
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1539..1558)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1593..1612
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1843..2200
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1843..2034
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1866..1886)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2051..2186
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2113..2132
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2231..3268
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3247..3255
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    3292..3340
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3496..3513)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3667..4255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3747..3766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4455..5246,5247..5315))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    5420..5468
                     /note="synthetic polyadenylation signal"
     misc_feature    5482..5573
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5522..5541
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct ctcgggtcct ctgagggacc tggggagctc aggctgggaa
       61 tctccaaggc agtaggtcgc ctatcaaaaa tcaaagtcca ggtttgtggg gggaaaacaa
      121 aagcagccca ttacccagag gactgtccgc cttcccctca ccccagccta ggcctttgaa
      181 aggaaacaaa agacaagaca aaatgattgg cgtcctgagg gagattcagc ctagagctct
      241 ctctccccca atccctccct ccggctgagg aaactaacaa aggaaaaaaa aattgcggaa
      301 agcaggattt agaggaagca aattccactg gtgcccttgg ctgccgggaa cgtggactag
      361 agagtctgcg gcgcagcccc gagcccagcg cttcccgcgc gtcttaggcc ggcgggcccg
      421 ggcgggggaa ggggacgcag accgcggacc ctaagacacc tgctgtaccc tccaccccca
      481 ccccacccac ctccccccaa ctccctagat gtgtcgtggg cggctgaacg tcgcccgttt
      541 aaggggcggg ccccggctcc acgtgctttc tgctgagtga ctgaactaca taaacagagg
      601 ccgggaacgg ggcggggagg agggagagca caggctttga ccgatagtaa cctctgcgct
      661 cggtgcagcc gaatctataa aaggaactag tcccggcaaa aaccccgtaa ttgcgagcga
      721 gagtgagtgg ggccgggacc cgcagagccg agccgaccct tctctcccgg gctgcggcag
      781 ggcagggcgg ggagctccgc gcaccaagct tggcaatccg gtactgttgg taaagccacc
      841 atggtcttca cactcgaaga tttcgttggg gactggcgac agacagccgg ctacaacctg
      901 gaccaagtcc ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg ggtgtccgta
      961 actccgatcc aaaggattgt cctgagcggt gaaaatgggc tgaagatcga catccatgtc
     1021 atcatcccgt atgaaggtct gagcggcgac caaatgggcc agatcgaaaa aatttttaag
     1081 gtggtgtacc ctgtggatga tcatcacttt aaggtgatcc tgcactatgg cacactggta
     1141 atcgacgggg ttacgccgaa catgatcgac tatttcggac ggccgtatga aggcatcgcc
     1201 gtgttcgacg gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa caaaattatc
     1261 gacgagcgcc tgatcaaccc cgacggctcc ctgctgttcc gagtaaccat caacggagtg
     1321 accggctggc ggctgtgcga acgcattctg gcgaattctc acggcttccc tcccgaggtg
     1381 gaggagcagg ccgccggcac cctgcccatg agctgcgccc aggagagcgg catggataga
     1441 caccctgctg cttgcgccag cgccaggatc aacgtctaag gccgcgactc tagagtcggg
     1501 gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     1561 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     1621 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     1681 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     1741 aaatcgataa ggatccgttt gcgtattggg cgctcttccg ctgatctgcg cagcaccatg
     1801 gcctgaaata acctctgaaa gaggaacttg gttagctacc ttctgaggcg gaaagaacca
     1861 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     1921 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     1981 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2041 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2101 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     2161 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcgatt cttctgacac
     2221 tagcgccacc atgaagaagc ccgaactcac cgctaccagc gttgaaaaat ttctcatcga
     2281 gaagttcgac agtgtgagcg acctgatgca gttgtcggag ggcgaagaga gccgagcctt
     2341 cagcttcgat gtcggcggac gcggctatgt actgcgggtg aatagctgcg ctgatggctt
     2401 ctacaaagac cgctacgtgt accgccactt cgccagcgct gcactaccca tccccgaagt
     2461 gttggacatc ggcgagttca gcgagagcct gacatactgc atcagtagac gcgcccaagg
     2521 cgttactctc caagacctcc ccgaaacaga gctgcctgct gtgttacagc ctgtcgccga
     2581 agctatggat gctattgccg ccgccgacct cagtcaaacc agcggcttcg gcccattcgg
     2641 gccccaaggc atcggccagt acacaacctg gcgggatttc atttgcgcca ttgctgatcc
     2701 ccatgtctac cactggcaga ccgtgatgga cgacaccgtg tccgccagcg tagctcaagc
     2761 cctggacgaa ctgatgctgt gggccgaaga ctgtcccgag gtgcgccacc tcgtccatgc
     2821 cgacttcggc agcaacaacg tcctgaccga caacggccgc atcaccgccg taatcgactg
     2881 gtccgaagct atgttcgggg acagtcagta cgaggtggcc aacatcttct tctggcggcc
     2941 ctggctggct tgcatggagc agcagactcg ctacttcgag cgccggcatc ccgagctggc
     3001 cggcagccct cgtctgcgag cctacatgct gcgcatcggc ctggatcagc tctaccagag
     3061 cctcgtggac ggcaacttcg acgatgctgc ctgggctcaa ggccgctgcg atgccatcgt
     3121 ccgcagcggg gccggcaccg tcggtcgcac acaaatcgct cgccggagcg cagccgtatg
     3181 gaccgacggc tgcgtcgagg tgctggccga cagcggcaac cgccggccca gtacacgacc
     3241 gcgcgctaag gaggtaggtc gagtttaaac tctagaaccg gtcatggccg caataaaata
     3301 tctttatttt cattacatct gtgtgttggt tttttgtgtg ttcgaactag atgctgtcga
     3361 ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact
     3421 atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca
     3481 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3541 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3601 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3661 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3721 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3781 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3841 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3901 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3961 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4021 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4081 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4141 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4201 cggtggtttt tttgtttgca agcagcagat tacgcacaga aaaaaaggat ctcaagaaga
     4261 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4321 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4381 ttttaaatca atctaaagta tatatgagta aacttggtct gacagcggcc gcaaatgcta
     4441 aaccactgca gtggttacca gtgcttgatc agtgaggcac cgatctcagc gatctgccta
     4501 tttcgttcgt ccatagtggc ctgactcccc gtcgtgtaga tcactacgat tcgtgagggc
     4561 ttaccatcag gccccagcgc agcaatgatg ccgcgagagc cgcgttcacc ggcccccgat
     4621 ttgtcagcaa tgaaccagcc agcagggagg gccgagcgaa gaagtggtcc tgctactttg
     4681 tccgcctcca tccagtctat gagctgctgt cgtgatgcta gagtaagaag ttcgccagtg
     4741 agtagtttcc gaagagttgt ggccattgct actggcatcg tggtatcacg ctcgtcgttc
     4801 ggtatggctt cgttcaactc tggttcccag cggtcaagcc gggtcacatg atcacccata
     4861 ttatgaagaa atgcagtcag ctccttaggg cctccgatcg ttgtcagaag taagttggcc
     4921 gcggtgttgt cgctcatggt aatggcagca ctacacaatt ctcttaccgt catgccatcc
     4981 gtaagatgct tttccgtgac cggcgagtac tcaaccaagt cgttttgtga gtagtgtata
     5041 cggcgaccaa gctgctcttg cccggcgtct atacgggaca acaccgcgcc acatagcagt
     5101 actttgaaag tgctcatcat cgggaatcgt tcttcggggc ggaaagactc aaggatcttg
     5161 ccgctattga gatccagttc gatatagccc actcttgcac ccagttgatc ttcagcatct
     5221 tttactttca ccagcgtttc ggggtgtgca aaaacaggca agcaaaatgc cgcaaagaag
     5281 ggaatgagtg cgacacgaaa atgttggatg ctcatactcg tcctttttca atattattga
     5341 agcatttatc agggttacta gtacgtctct caaggataag taagtaatat taaggtacgg
     5401 gaggtattgg acaggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt
     5461 tttgtgtgaa tcgatagtac taacatacgc tctccatcaa aacaaaacga aacaaaacaa
     5521 actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc tct
//
