LOCUS       sequence-471829-        6432 bp    DNA     circular SYN 28-MAY-2025
DEFINITION  Express 2B6 response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6432)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6432)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 28, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGL4 [luc2P/2B6/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..6432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      239..248
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        499..530
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      557..566
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             563..2212
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             2216..2335
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2387..2508
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2398..2417)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2452..2471
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2702..3059
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2702..2893
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2725..2745)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2910..3045
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2972..2991
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3090..4127
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4106..4114
                     /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    4151..4199
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4355..4372)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4526..5114)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4606..4625)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5314..6105,6106..6174))
                     /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    6279..6327
                     /note="synthetic polyadenylation signal"
     misc_feature    6341..6432
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6381..6400
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtaccc tttctccatc cacaaaatgg gggtgaaaac aattactaca
       61 tcaaagggtg acaactaagt gaattactct attgaaagca cttcatgcct ccttatcagt
      121 gctccccaag ggtggaatga ggacccagag tacccgtatt aggccatact tgtgttgcta
      181 taaagactgt gccagattgc acaacacagc aggagacagg tcctggggtc aggaaagtcc
      241 atggtggcat ctttcctagg tcaggaaagt gtgggttgta ctttgttggt cctctcctcg
      301 tgggttgggg aagtatgggg tgcagctgct cctggcagct ttgactttca gcctcagata
      361 atgcattgcc tgtgtctaat ccacatggat gtggggtgac tcagccacct ccattccctg
      421 aatcagcatc ccttggttca ggaaagtcca tgctgccacc tcttcagggt caggaaagta
      481 cagtttaagc ttagacacta gagggtatat aatggaagct cgacttccag cttggcaatc
      541 cggtactgtt ggtaaagcca ccatggaaga tgccaaaaac attaagaagg gcccagcgcc
      601 attctaccca ctcgaagacg ggaccgccgg cgagcagctg cacaaagcca tgaagcgcta
      661 cgccctggtg cccggcacca tcgcctttac cgacgcacat atcgaggtgg acattaccta
      721 cgccgagtac ttcgagatga gcgttcggct ggcagaagct atgaagcgct atgggctgaa
      781 tacaaaccat cggatcgtgg tgtgcagcga gaatagcttg cagttcttca tgcccgtgtt
      841 gggtgccctg ttcatcggtg tggctgtggc cccagctaac gacatctaca acgagcgcga
      901 gctgctgaac agcatgggca tcagccagcc caccgtcgta ttcgtgagca agaaagggct
      961 gcaaaagatc ctcaacgtgc aaaagaagct accgatcata caaaagatca tcatcatgga
     1021 tagcaagacc gactaccagg gcttccaaag catgtacacc ttcgtgactt cccatttgcc
     1081 acccggcttc aacgagtacg acttcgtgcc cgagagcttc gaccgggaca aaaccatcgc
     1141 cctgatcatg aacagtagtg gcagtaccgg attgcccaag ggcgtagccc taccgcaccg
     1201 caccgcttgt gtccgattca gtcatgcccg cgaccccatc ttcggcaacc agatcatccc
     1261 cgacaccgct atcctcagcg tggtgccatt tcaccacggc ttcggcatgt tcaccacgct
     1321 gggctacttg atctgcggct ttcgggtcgt gctcatgtac cgcttcgagg aggagctatt
     1381 cttgcgcagc ttgcaagact ataagattca atctgccctg ctggtgccca cactatttag
     1441 cttcttcgct aagagcactc tcatcgacaa gtacgaccta agcaacttgc acgagatcgc
     1501 cagcggcggg gcgccgctca gcaaggaggt aggtgaggcc gtggccaaac gcttccacct
     1561 accaggcatc cgccagggct acggcctgac agaaacaacc agcgccattc tgatcacccc
     1621 cgaaggggac gacaagcctg gcgcagtagg caaggtggtg cccttcttcg aggctaaggt
     1681 ggtggacttg gacaccggta agacactggg tgtgaaccag cgcggcgagc tgtgcgtccg
     1741 tggccccatg atcatgagcg gctacgttaa caaccccgag gctacaaacg ctctcatcga
     1801 caaggacggc tggctgcaca gcggcgacat cgcctactgg gacgaggacg agcacttctt
     1861 catcgtggac cggctgaaga gcctgatcaa atacaagggc taccaggtag ccccagccga
     1921 actggagagc atcctgctgc aacaccccaa catcttcgac gccggggtcg ccggcctgcc
     1981 cgacgacgat gccggcgagc tgcccgccgc agtcgtcgtg ctggaacacg gtaaaaccat
     2041 gaccgagaag gagatcgtgg actatgtggc cagccaggtt acaaccgcca agaagctgcg
     2101 cggtggtgtt gtgttcgtgg acgaggtgcc taaaggactg accggcaagt tggacgcccg
     2161 caagatccgc gagattctca ttaaggccaa gaagggcggc aagatcgccg tgaattctca
     2221 cggcttccct cccgaggtgg aggagcaggc cgccggcacc ctgcccatga gctgcgccca
     2281 ggagagcggc atggatagac accctgctgc ttgcgccagc gccaggatca acgtctaagg
     2341 ccgcgactct agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat
     2401 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     2461 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     2521 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     2581 aacctctaca aatgtggtaa aatcgataag gatccgtttg cgtattgggc gctcttccgc
     2641 tgatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttagctacct
     2701 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     2761 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     2821 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2881 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2941 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3001 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3061 agctcgattc ttctgacact agcgccacca tgaagaagcc cgaactcacc gctaccagcg
     3121 ttgaaaaatt tctcatcgag aagttcgaca gtgtgagcga cctgatgcag ttgtcggagg
     3181 gcgaagagag ccgagccttc agcttcgatg tcggcggacg cggctatgta ctgcgggtga
     3241 atagctgcgc tgatggcttc tacaaagacc gctacgtgta ccgccacttc gccagcgctg
     3301 cactacccat ccccgaagtg ttggacatcg gcgagttcag cgagagcctg acatactgca
     3361 tcagtagacg cgcccaaggc gttactctcc aagacctccc cgaaacagag ctgcctgctg
     3421 tgttacagcc tgtcgccgaa gctatggatg ctattgccgc cgccgacctc agtcaaacca
     3481 gcggcttcgg cccattcggg ccccaaggca tcggccagta cacaacctgg cgggatttca
     3541 tttgcgccat tgctgatccc catgtctacc actggcagac cgtgatggac gacaccgtgt
     3601 ccgccagcgt agctcaagcc ctggacgaac tgatgctgtg ggccgaagac tgtcccgagg
     3661 tgcgccacct cgtccatgcc gacttcggca gcaacaacgt cctgaccgac aacggccgca
     3721 tcaccgccgt aatcgactgg tccgaagcta tgttcgggga cagtcagtac gaggtggcca
     3781 acatcttctt ctggcggccc tggctggctt gcatggagca gcagactcgc tacttcgagc
     3841 gccggcatcc cgagctggcc ggcagccctc gtctgcgagc ctacatgctg cgcatcggcc
     3901 tggatcagct ctaccagagc ctcgtggacg gcaacttcga cgatgctgcc tgggctcaag
     3961 gccgctgcga tgccatcgtc cgcagcgggg ccggcaccgt cggtcgcaca caaatcgctc
     4021 gccggagcgc agccgtatgg accgacggct gcgtcgaggt gctggccgac agcggcaacc
     4081 gccggcccag tacacgaccg cgcgctaagg aggtaggtcg agtttaaact ctagaaccgg
     4141 tcatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtgt
     4201 tcgaactaga tgctgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg
     4261 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
     4321 cgtaggacag gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4381 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4441 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4501 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4561 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4621 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4681 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4741 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4801 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4861 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4921 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4981 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5041 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5101 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5161 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5221 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5281 acagcggccg caaatgctaa accactgcag tggttaccag tgcttgatca gtgaggcacc
     5341 gatctcagcg atctgcctat ttcgttcgtc catagtggcc tgactccccg tcgtgtagat
     5401 cactacgatt cgtgagggct taccatcagg ccccagcgca gcaatgatgc cgcgagagcc
     5461 gcgttcaccg gcccccgatt tgtcagcaat gaaccagcca gcagggaggg ccgagcgaag
     5521 aagtggtcct gctactttgt ccgcctccat ccagtctatg agctgctgtc gtgatgctag
     5581 agtaagaagt tcgccagtga gtagtttccg aagagttgtg gccattgcta ctggcatcgt
     5641 ggtatcacgc tcgtcgttcg gtatggcttc gttcaactct ggttcccagc ggtcaagccg
     5701 ggtcacatga tcacccatat tatgaagaaa tgcagtcagc tccttagggc ctccgatcgt
     5761 tgtcagaagt aagttggccg cggtgttgtc gctcatggta atggcagcac tacacaattc
     5821 tcttaccgtc atgccatccg taagatgctt ttccgtgacc ggcgagtact caaccaagtc
     5881 gttttgtgag tagtgtatac ggcgaccaag ctgctcttgc ccggcgtcta tacgggacaa
     5941 caccgcgcca catagcagta ctttgaaagt gctcatcatc gggaatcgtt cttcggggcg
     6001 gaaagactca aggatcttgc cgctattgag atccagttcg atatagccca ctcttgcacc
     6061 cagttgatct tcagcatctt ttactttcac cagcgtttcg gggtgtgcaa aaacaggcaa
     6121 gcaaaatgcc gcaaagaagg gaatgagtgc gacacgaaaa tgttggatgc tcatactcgt
     6181 cctttttcaa tattattgaa gcatttatca gggttactag tacgtctctc aaggataagt
     6241 aagtaatatt aaggtacggg aggtattgga caggccgcaa taaaatatct ttattttcat
     6301 tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa
     6361 acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca
     6421 gaacatttct ct
//
