LOCUS       sequence-471825-        8007 bp    DNA     circular SYN 18-JUN-2025
DEFINITION  Express PSA-long response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8007)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 8007)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 18, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGL4 [luc2P/PSA-long/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..8007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2132..2141
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2138..3787
                     /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             3791..3910
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    3962..4083
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3973..3992)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4027..4046
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4277..4634
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4277..4468
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4300..4320)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4485..4620
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4547..4566
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4665..5702
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5681..5689
                     /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    5726..5774
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5930..5947)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6101..6689)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6181..6200)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6889..7680,7681..7749))
                     /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    7854..7902
                     /note="synthetic polyadenylation signal"
     misc_feature    7916..8007
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7956..7975
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtaccg aattctctag aaatctagct gatatagtgt ggctcaaaac
       61 cttcagcaca aatcacaccg ttagactatc tggtgtggcc caaaccttca ggtgaacaaa
      121 ggcactctaa actggcagga tattccaaag cattagagat gacctcttgc aaagaaaaag
      181 aaatggaaaa gaaaaagaaa gaaaggaaaa aaaaaaaaaa aaagagatga cctctcaggc
      241 tctgagggga aacgcctgag gtctttgagc aaggtcagtc ctctgttgca cagtctccct
      301 cacagggtca ttgtgacgat caaatgtggt cacgtgtatg aggcaccagc acatgcctgg
      361 ctctggggag tgccgtgtaa gtgtatgctt gcactgctga atggctggga tgtgtcaggg
      421 attatcttca gcacttacag atgctcatct catcctcaca gcatcactat gggatgggta
      481 ttactggcct catttgatgg agaaactggc tgtggctcag aaagggggga ccactagacc
      541 agggacactc tggatgctgg ggactccaga gaccatgacc actcaccaac tgcagagaaa
      601 ttaattgtgg cctgatgtcc ctgtcctgga gagggtggag gtggaccttc actaacctcc
      661 taccttgacc ctctctttta gggctctttc tgacctccac catgatacta ggaccccatt
      721 gtattctgta ccctcttgac tctatgaccc ccactgccca ctgcatccag ctgggtcccc
      781 tcctatctct attcccagct ggccagtgca gtctcagtgc ccacctgttt gtcagtaact
      841 ctgaaggggc tgacatttta ctgacttgca aacaaataag ctaactttcc agagttttgt
      901 gaatgctggc agagtccatg agactcctga gtcagaggca aaggctttta ctgctcacag
      961 cttagcagac agcatgaggt tcatgttcac attagtacac cttgcccccc ccaaatcttg
     1021 tagggtgacc agagcagtct aggtggatgc tgtgcacacg gggtttgtgc cactggtgag
     1081 aaacctgaga ttaggaatcc tcaatcttat actgggacaa cttgcaaacc tgctcagcct
     1141 ttgtctctga tgaagatatt atcttcatga tcttggattg aaaacagacc tactctggag
     1201 gaacatattg tatcgattgt ccttgacagt aaacaaatct gttgtaagag acattatctt
     1261 tattatctag gacagtaagc aagcctggat ctgagagaga tatcatcttg caaggatgcc
     1321 tgctttacaa acatccttga aacaacaatc cagaaaaaaa aaggtgttgc tgtctttgct
     1381 cagaagacac acagatacgt gacagaacca tggagaattg cctcccaaca ctgttcagcc
     1441 agagccttcc acccttgtct gcaggaattc cacattgttt gctgcacgtt ggattttgaa
     1501 atgctaggga actttgggag actcatattt ctgggctaga ggatctgtgg accacaagat
     1561 ctttttatga tgacagtagc aatgtatctg tggagctgga ttctgggttg ggagtgcaag
     1621 gaaaagaatg tactaaatgc caagacatct atttcaggag catgaggaat aaaagttcta
     1681 gtttctggtc tcagagtggt gcagggatca gggagtctca caatctcctg agtgctggtg
     1741 tcttagggca cactgggtct tggagtgcaa aggatctagg cacgtgaggc tttgtatgaa
     1801 gaatcgggga tcgtacccac cccctgtttc tgtttcatcc tgggcgtgtc tcctctgcct
     1861 ttgtccccta gatgaagtct ccatgagcta cagggcctgg tgcatccagg gtgatctagt
     1921 aattgcagaa cagcaagtgc tagctctccc tccccttcca cagctctggg tgtgggaggg
     1981 ggttgtccag cctccagcag catggggagg gccttggtca gcctctgggt gccagcaggg
     2041 caggggcgga gtcctgggga atgaaggttt tatagggctc ctgggggagg ctccccagcc
     2101 ccaagcttgg caatccggta ctgttggtaa agccaccatg gaagatgcca aaaacattaa
     2161 gaagggccca gcgccattct acccactcga agacgggacc gccggcgagc agctgcacaa
     2221 agccatgaag cgctacgccc tggtgcccgg caccatcgcc tttaccgacg cacatatcga
     2281 ggtggacatt acctacgccg agtacttcga gatgagcgtt cggctggcag aagctatgaa
     2341 gcgctatggg ctgaatacaa accatcggat cgtggtgtgc agcgagaata gcttgcagtt
     2401 cttcatgccc gtgttgggtg ccctgttcat cggtgtggct gtggccccag ctaacgacat
     2461 ctacaacgag cgcgagctgc tgaacagcat gggcatcagc cagcccaccg tcgtattcgt
     2521 gagcaagaaa gggctgcaaa agatcctcaa cgtgcaaaag aagctaccga tcatacaaaa
     2581 gatcatcatc atggatagca agaccgacta ccagggcttc caaagcatgt acaccttcgt
     2641 gacttcccat ttgccacccg gcttcaacga gtacgacttc gtgcccgaga gcttcgaccg
     2701 ggacaaaacc atcgccctga tcatgaacag tagtggcagt accggattgc ccaagggcgt
     2761 agccctaccg caccgcaccg cttgtgtccg attcagtcat gcccgcgacc ccatcttcgg
     2821 caaccagatc atccccgaca ccgctatcct cagcgtggtg ccatttcacc acggcttcgg
     2881 catgttcacc acgctgggct acttgatctg cggctttcgg gtcgtgctca tgtaccgctt
     2941 cgaggaggag ctattcttgc gcagcttgca agactataag attcaatctg ccctgctggt
     3001 gcccacacta tttagcttct tcgctaagag cactctcatc gacaagtacg acctaagcaa
     3061 cttgcacgag atcgccagcg gcggggcgcc gctcagcaag gaggtaggtg aggccgtggc
     3121 caaacgcttc cacctaccag gcatccgcca gggctacggc ctgacagaaa caaccagcgc
     3181 cattctgatc acccccgaag gggacgacaa gcctggcgca gtaggcaagg tggtgccctt
     3241 cttcgaggct aaggtggtgg acttggacac cggtaagaca ctgggtgtga accagcgcgg
     3301 cgagctgtgc gtccgtggcc ccatgatcat gagcggctac gttaacaacc ccgaggctac
     3361 aaacgctctc atcgacaagg acggctggct gcacagcggc gacatcgcct actgggacga
     3421 ggacgagcac ttcttcatcg tggaccggct gaagagcctg atcaaataca agggctacca
     3481 ggtagcccca gccgaactgg agagcatcct gctgcaacac cccaacatct tcgacgccgg
     3541 ggtcgccggc ctgcccgacg acgatgccgg cgagctgccc gccgcagtcg tcgtgctgga
     3601 acacggtaaa accatgaccg agaaggagat cgtggactat gtggccagcc aggttacaac
     3661 cgccaagaag ctgcgcggtg gtgttgtgtt cgtggacgag gtgcctaaag gactgaccgg
     3721 caagttggac gcccgcaaga tccgcgagat tctcattaag gccaagaagg gcggcaagat
     3781 cgccgtgaat tctcacggct tccctcccga ggtggaggag caggccgccg gcaccctgcc
     3841 catgagctgc gcccaggaga gcggcatgga tagacaccct gctgcttgcg ccagcgccag
     3901 gatcaacgtc taaggccgcg actctagagt cggggcggcc ggccgcttcg agcagacatg
     3961 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     4021 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     4081 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     4141 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gtttgcgtat
     4201 tgggcgctct tccgctgatc tgcgcagcac catggcctga aataacctct gaaagaggaa
     4261 cttggttagc taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     4321 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4381 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4441 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4501 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4561 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4621 taggcttttg caaaaagctc gattcttctg acactagcgc caccatgaag aagcccgaac
     4681 tcaccgctac cagcgttgaa aaatttctca tcgagaagtt cgacagtgtg agcgacctga
     4741 tgcagttgtc ggagggcgaa gagagccgag ccttcagctt cgatgtcggc ggacgcggct
     4801 atgtactgcg ggtgaatagc tgcgctgatg gcttctacaa agaccgctac gtgtaccgcc
     4861 acttcgccag cgctgcacta cccatccccg aagtgttgga catcggcgag ttcagcgaga
     4921 gcctgacata ctgcatcagt agacgcgccc aaggcgttac tctccaagac ctccccgaaa
     4981 cagagctgcc tgctgtgtta cagcctgtcg ccgaagctat ggatgctatt gccgccgccg
     5041 acctcagtca aaccagcggc ttcggcccat tcgggcccca aggcatcggc cagtacacaa
     5101 cctggcggga tttcatttgc gccattgctg atccccatgt ctaccactgg cagaccgtga
     5161 tggacgacac cgtgtccgcc agcgtagctc aagccctgga cgaactgatg ctgtgggccg
     5221 aagactgtcc cgaggtgcgc cacctcgtcc atgccgactt cggcagcaac aacgtcctga
     5281 ccgacaacgg ccgcatcacc gccgtaatcg actggtccga agctatgttc ggggacagtc
     5341 agtacgaggt ggccaacatc ttcttctggc ggccctggct ggcttgcatg gagcagcaga
     5401 ctcgctactt cgagcgccgg catcccgagc tggccggcag ccctcgtctg cgagcctaca
     5461 tgctgcgcat cggcctggat cagctctacc agagcctcgt ggacggcaac ttcgacgatg
     5521 ctgcctgggc tcaaggccgc tgcgatgcca tcgtccgcag cggggccggc accgtcggtc
     5581 gcacacaaat cgctcgccgg agcgcagccg tatggaccga cggctgcgtc gaggtgctgg
     5641 ccgacagcgg caaccgccgg cccagtacac gaccgcgcgc taaggaggta ggtcgagttt
     5701 aaactctaga accggtcatg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     5761 tggttttttg tgtgttcgaa ctagatgctg tcgaccgatg cccttgagag ccttcaaccc
     5821 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
     5881 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg
     5941 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6001 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6061 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6121 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6181 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6241 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6301 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6361 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6421 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6481 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6541 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6601 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6661 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6721 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6781 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6841 agtaaacttg gtctgacagc ggccgcaaat gctaaaccac tgcagtggtt accagtgctt
     6901 gatcagtgag gcaccgatct cagcgatctg cctatttcgt tcgtccatag tggcctgact
     6961 ccccgtcgtg tagatcacta cgattcgtga gggcttacca tcaggcccca gcgcagcaat
     7021 gatgccgcga gagccgcgtt caccggcccc cgatttgtca gcaatgaacc agccagcagg
     7081 gagggccgag cgaagaagtg gtcctgctac tttgtccgcc tccatccagt ctatgagctg
     7141 ctgtcgtgat gctagagtaa gaagttcgcc agtgagtagt ttccgaagag ttgtggccat
     7201 tgctactggc atcgtggtat cacgctcgtc gttcggtatg gcttcgttca actctggttc
     7261 ccagcggtca agccgggtca catgatcacc catattatga agaaatgcag tcagctcctt
     7321 agggcctccg atcgttgtca gaagtaagtt ggccgcggtg ttgtcgctca tggtaatggc
     7381 agcactacac aattctctta ccgtcatgcc atccgtaaga tgcttttccg tgaccggcga
     7441 gtactcaacc aagtcgtttt gtgagtagtg tatacggcga ccaagctgct cttgcccggc
     7501 gtctatacgg gacaacaccg cgccacatag cagtactttg aaagtgctca tcatcgggaa
     7561 tcgttcttcg gggcggaaag actcaaggat cttgccgcta ttgagatcca gttcgatata
     7621 gcccactctt gcacccagtt gatcttcagc atcttttact ttcaccagcg tttcggggtg
     7681 tgcaaaaaca ggcaagcaaa atgccgcaaa gaagggaatg agtgcgacac gaaaatgttg
     7741 gatgctcata ctcgtccttt ttcaatatta ttgaagcatt tatcagggtt actagtacgt
     7801 ctctcaagga taagtaagta atattaaggt acgggaggta ttggacaggc cgcaataaaa
     7861 tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gtactaacat
     7921 acgctctcca tcaaaacaaa acgaaacaaa acaaactagc aaaataggct gtccccagtg
     7981 caagtgcagg tgccagaaca tttctct
//
