LOCUS       sequence-470901-        4923 bp    DNA     circular SYN 02-MAY-2025
DEFINITION  Express GAS response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4923)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 4923)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pNL [NlucP/GAS-RE/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..4923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        127..158
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      185..194
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             191..703
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             707..826
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    878..999
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(889..908)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     943..962
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1193..1550
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1193..1384
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1216..1236)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1401..1536
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1463..1482
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1581..2618
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2597..2605
                     /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    2642..2690
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2846..2863)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3017..3605)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3097..3116)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3805..4596,4597..4665))
                     /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    4770..4818
                     /note="synthetic polyadenylation signal"
     misc_feature    4832..4923
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4872..4891
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcagtt tcatattact ctaaatcagt ttcatattac
       61 tctaaatcag tttcatatta ctctaaatca gtttcatatt actctaaatc ctcgaagctt
      121 agacactaga gggtatataa tggaagctcg acttccagct tggcaatccg gtactgttgg
      181 taaagccacc atggtcttca cactcgaaga tttcgttggg gactggcgac agacagccgg
      241 ctacaacctg gaccaagtcc ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg
      301 ggtgtccgta actccgatcc aaaggattgt cctgagcggt gaaaatgggc tgaagatcga
      361 catccatgtc atcatcccgt atgaaggtct gagcggcgac caaatgggcc agatcgaaaa
      421 aatttttaag gtggtgtacc ctgtggatga tcatcacttt aaggtgatcc tgcactatgg
      481 cacactggta atcgacgggg ttacgccgaa catgatcgac tatttcggac ggccgtatga
      541 aggcatcgcc gtgttcgacg gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa
      601 caaaattatc gacgagcgcc tgatcaaccc cgacggctcc ctgctgttcc gagtaaccat
      661 caacggagtg accggctggc ggctgtgcga acgcattctg gcgaattctc acggctttcc
      721 gcctgaggtt gaagagcaag ccgccggtac attgcctatg tcctgcgcac aagaaagcgg
      781 tatggaccgg cacccagccg cttgtgcttc agctcgcatc aacgtctaag gccgcgactc
      841 tagagtcggg gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga
      901 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
      961 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     1021 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     1081 aaatgtggta aaatcgataa ggatccgttt gcgtattggg cgctcttccg ctgatctgcg
     1141 cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttagctacc ttctgaggcg
     1201 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     1261 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     1321 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     1381 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     1441 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     1501 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcgatt
     1561 cttctgacac tagcgccacc atgaagaagc ccgaactcac cgctaccagc gttgaaaaat
     1621 ttctcatcga gaagttcgac agtgtgagcg acctgatgca gttgtcggag ggcgaagaga
     1681 gccgagcctt cagcttcgat gtcggcggac gcggctatgt actgcgggtg aatagctgcg
     1741 ctgatggctt ctacaaagac cgctacgtgt accgccactt cgccagcgct gcactaccca
     1801 tccccgaagt gttggacatc ggcgagttca gcgagagcct gacatactgc atcagtagac
     1861 gcgcccaagg cgttactctc caagacctcc ccgaaacaga gctgcctgct gtgttacagc
     1921 ctgtcgccga agctatggat gctattgccg ccgccgacct cagtcaaacc agcggcttcg
     1981 gcccattcgg gccccaaggc atcggccagt acacaacctg gcgggatttc atttgcgcca
     2041 ttgctgatcc ccatgtctac cactggcaga ccgtgatgga cgacaccgtg tccgccagcg
     2101 tagctcaagc cctggacgaa ctgatgctgt gggccgaaga ctgtcccgag gtgcgccacc
     2161 tcgtccatgc cgacttcggc agcaacaacg tcctgaccga caacggccgc atcaccgccg
     2221 taatcgactg gtccgaagct atgttcgggg acagtcagta cgaggtggcc aacatcttct
     2281 tctggcggcc ctggctggct tgcatggagc agcagactcg ctacttcgag cgccggcatc
     2341 ccgagctggc cggcagccct cgtctgcgag cctacatgct gcgcatcggc ctggatcagc
     2401 tctaccagag cctcgtggac ggcaacttcg acgatgctgc ctgggctcaa ggccgctgcg
     2461 atgccatcgt ccgcagcggg gccggcaccg tcggtcgcac acaaatcgct cgccggagcg
     2521 cagccgtatg gaccgacggc tgcgtcgagg tgctggccga cagcggcaac cgccggccca
     2581 gtacacgacc gcgcgctaag gaggtaggtc gagtttaaac tctagaaccg gtcatggccg
     2641 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg ttcgaactag
     2701 atgctgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
     2761 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
     2821 ggtgccggca gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2881 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2941 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3001 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3061 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3121 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3181 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3241 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3301 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3361 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3421 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3481 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3541 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3601 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3661 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3721 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagcggcc
     3781 gcaaatgcta aaccactgca gtggttacca gtgcttgatc agtgaggcac cgatctcagc
     3841 gatctgccta tttcgttcgt ccatagtggc ctgactcccc gtcgtgtaga tcactacgat
     3901 tcgtgagggc ttaccatcag gccccagcgc agcaatgatg ccgcgagagc cgcgttcacc
     3961 ggcccccgat ttgtcagcaa tgaaccagcc agcagggagg gccgagcgaa gaagtggtcc
     4021 tgctactttg tccgcctcca tccagtctat gagctgctgt cgtgatgcta gagtaagaag
     4081 ttcgccagtg agtagtttcc gaagagttgt ggccattgct actggcatcg tggtatcacg
     4141 ctcgtcgttc ggtatggctt cgttcaactc tggttcccag cggtcaagcc gggtcacatg
     4201 atcacccata ttatgaagaa atgcagtcag ctccttaggg cctccgatcg ttgtcagaag
     4261 taagttggcc gcggtgttgt cgctcatggt aatggcagca ctacacaatt ctcttaccgt
     4321 catgccatcc gtaagatgct tttccgtgac cggcgagtac tcaaccaagt cgttttgtga
     4381 gtagtgtata cggcgaccaa gctgctcttg cccggcgtct atacgggaca acaccgcgcc
     4441 acatagcagt actttgaaag tgctcatcat cgggaatcgt tcttcggggc ggaaagactc
     4501 aaggatcttg ccgctattga gatccagttc gatatagccc actcttgcac ccagttgatc
     4561 ttcagcatct tttactttca ccagcgtttc ggggtgtgca aaaacaggca agcaaaatgc
     4621 cgcaaagaag ggaatgagtg cgacacgaaa atgttggatg ctcatactcg tcctttttca
     4681 atattattga agcatttatc agggttacta gtacgtctct caaggataag taagtaatat
     4741 taaggtacgg gaggtattgg acaggccgca ataaaatatc tttattttca ttacatctgt
     4801 gtgttggttt tttgtgtgaa tcgatagtac taacatacgc tctccatcaa aacaaaacga
     4861 aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc
     4921 tct
//
