LOCUS       sequence-472952-        6252 bp    DNA     circular SYN 15-MAY-2025
DEFINITION  Express GCSF response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6252)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6252)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGL4 [luc2P/GCSF/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..6252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    600..648
                     /note="synthetic polyadenylation signal"
     misc_feature    662..753
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     702..721
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      1130..1139
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1136..2785
                     /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             2789..2908
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2960..3081
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2971..2990)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3025..3044
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3275..3632
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3275..3466
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3298..3318)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3483..3618
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3545..3564
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3663..4700
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4679..4687
                     /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    4724..4772
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4928..4945)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5099..5687)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5179..5198)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5887..6252,1..426,427..495))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 tggttcccag cggtcaagcc gggtcacatg atcacccata ttatgaagaa atgcagtcag
       61 ctccttaggg cctccgatcg ttgtcagaag taagttggcc gcggtgttgt cgctcatggt
      121 aatggcagca ctacacaatt ctcttaccgt catgccatcc gtaagatgct tttccgtgac
      181 cggcgagtac tcaaccaagt cgttttgtga gtagtgtata cggcgaccaa gctgctcttg
      241 cccggcgtct atacgggaca acaccgcgcc acatagcagt actttgaaag tgctcatcat
      301 cgggaatcgt tcttcggggc ggaaagactc aaggatcttg ccgctattga gatccagttc
      361 gatatagccc actcttgcac ccagttgatc ttcagcatct tttactttca ccagcgtttc
      421 ggggtgtgca aaaacaggca agcaaaatgc cgcaaagaag ggaatgagtg cgacacgaaa
      481 atgttggatg ctcatactcg tcctttttca atattattga agcatttatc agggttacta
      541 gtacgtctct caaggataag taagtaatat taaggtacgg gaggtattgg acaggccgca
      601 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac
      661 taacatacgc tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc
      721 ccagtgcaag tgcaggtgcc agaacatttc tctggcctaa ctggccggta ccaggcgtct
      781 atcagcggct cagcctttgt tcagctgttc tgttcaaaca ctctggggcc attcaggcct
      841 gggtggggca gcgggaggaa gggagtttga ggggggcaag gcgacgtcaa aggaggatca
      901 gagattccac aatttcacaa aactttcgca aacagctttt tgttccaacc cccctgcatt
      961 gtcttggaca ccaaatttgc ataaatcctg ggaagttatt actaagcctt agtcgtggcc
     1021 ccaggtaatt tcctcccagg cctccatggg gttatgtata aaggcccccc tagagctggg
     1081 ccccaaaaca gcccggagcc aagcttggca atccggtact gttggtaaag ccaccatgga
     1141 agatgccaaa aacattaaga agggcccagc gccattctac ccactcgaag acgggaccgc
     1201 cggcgagcag ctgcacaaag ccatgaagcg ctacgccctg gtgcccggca ccatcgcctt
     1261 taccgacgca catatcgagg tggacattac ctacgccgag tacttcgaga tgagcgttcg
     1321 gctggcagaa gctatgaagc gctatgggct gaatacaaac catcggatcg tggtgtgcag
     1381 cgagaatagc ttgcagttct tcatgcccgt gttgggtgcc ctgttcatcg gtgtggctgt
     1441 ggccccagct aacgacatct acaacgagcg cgagctgctg aacagcatgg gcatcagcca
     1501 gcccaccgtc gtattcgtga gcaagaaagg gctgcaaaag atcctcaacg tgcaaaagaa
     1561 gctaccgatc atacaaaaga tcatcatcat ggatagcaag accgactacc agggcttcca
     1621 aagcatgtac accttcgtga cttcccattt gccacccggc ttcaacgagt acgacttcgt
     1681 gcccgagagc ttcgaccggg acaaaaccat cgccctgatc atgaacagta gtggcagtac
     1741 cggattgccc aagggcgtag ccctaccgca ccgcaccgct tgtgtccgat tcagtcatgc
     1801 ccgcgacccc atcttcggca accagatcat ccccgacacc gctatcctca gcgtggtgcc
     1861 atttcaccac ggcttcggca tgttcaccac gctgggctac ttgatctgcg gctttcgggt
     1921 cgtgctcatg taccgcttcg aggaggagct attcttgcgc agcttgcaag actataagat
     1981 tcaatctgcc ctgctggtgc ccacactatt tagcttcttc gctaagagca ctctcatcga
     2041 caagtacgac ctaagcaact tgcacgagat cgccagcggc ggggcgccgc tcagcaagga
     2101 ggtaggtgag gccgtggcca aacgcttcca cctaccaggc atccgccagg gctacggcct
     2161 gacagaaaca accagcgcca ttctgatcac ccccgaaggg gacgacaagc ctggcgcagt
     2221 aggcaaggtg gtgcccttct tcgaggctaa ggtggtggac ttggacaccg gtaagacact
     2281 gggtgtgaac cagcgcggcg agctgtgcgt ccgtggcccc atgatcatga gcggctacgt
     2341 taacaacccc gaggctacaa acgctctcat cgacaaggac ggctggctgc acagcggcga
     2401 catcgcctac tgggacgagg acgagcactt cttcatcgtg gaccggctga agagcctgat
     2461 caaatacaag ggctaccagg tagccccagc cgaactggag agcatcctgc tgcaacaccc
     2521 caacatcttc gacgccgggg tcgccggcct gcccgacgac gatgccggcg agctgcccgc
     2581 cgcagtcgtc gtgctggaac acggtaaaac catgaccgag aaggagatcg tggactatgt
     2641 ggccagccag gttacaaccg ccaagaagct gcgcggtggt gttgtgttcg tggacgaggt
     2701 gcctaaagga ctgaccggca agttggacgc ccgcaagatc cgcgagattc tcattaaggc
     2761 caagaagggc ggcaagatcg ccgtgaattc tcacggcttc cctcccgagg tggaggagca
     2821 ggccgccggc accctgccca tgagctgcgc ccaggagagc ggcatggata gacaccctgc
     2881 tgcttgcgcc agcgccagga tcaacgtcta aggccgcgac tctagagtcg gggcggccgg
     2941 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     3001 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     3061 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     3121 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     3181 aaggatccgt ttgcgtattg ggcgctcttc cgctgatctg cgcagcacca tggcctgaaa
     3241 taacctctga aagaggaact tggttagcta ccttctgagg cggaaagaac cagctgtgga
     3301 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3361 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3421 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3481 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3541 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3601 gaggcttttt tggaggccta ggcttttgca aaaagctcga ttcttctgac actagcgcca
     3661 ccatgaagaa gcccgaactc accgctacca gcgttgaaaa atttctcatc gagaagttcg
     3721 acagtgtgag cgacctgatg cagttgtcgg agggcgaaga gagccgagcc ttcagcttcg
     3781 atgtcggcgg acgcggctat gtactgcggg tgaatagctg cgctgatggc ttctacaaag
     3841 accgctacgt gtaccgccac ttcgccagcg ctgcactacc catccccgaa gtgttggaca
     3901 tcggcgagtt cagcgagagc ctgacatact gcatcagtag acgcgcccaa ggcgttactc
     3961 tccaagacct ccccgaaaca gagctgcctg ctgtgttaca gcctgtcgcc gaagctatgg
     4021 atgctattgc cgccgccgac ctcagtcaaa ccagcggctt cggcccattc gggccccaag
     4081 gcatcggcca gtacacaacc tggcgggatt tcatttgcgc cattgctgat ccccatgtct
     4141 accactggca gaccgtgatg gacgacaccg tgtccgccag cgtagctcaa gccctggacg
     4201 aactgatgct gtgggccgaa gactgtcccg aggtgcgcca cctcgtccat gccgacttcg
     4261 gcagcaacaa cgtcctgacc gacaacggcc gcatcaccgc cgtaatcgac tggtccgaag
     4321 ctatgttcgg ggacagtcag tacgaggtgg ccaacatctt cttctggcgg ccctggctgg
     4381 cttgcatgga gcagcagact cgctacttcg agcgccggca tcccgagctg gccggcagcc
     4441 ctcgtctgcg agcctacatg ctgcgcatcg gcctggatca gctctaccag agcctcgtgg
     4501 acggcaactt cgacgatgct gcctgggctc aaggccgctg cgatgccatc gtccgcagcg
     4561 gggccggcac cgtcggtcgc acacaaatcg ctcgccggag cgcagccgta tggaccgacg
     4621 gctgcgtcga ggtgctggcc gacagcggca accgccggcc cagtacacga ccgcgcgcta
     4681 aggaggtagg tcgagtttaa actctagaac cggtcatggc cgcaataaaa tatctttatt
     4741 ttcattacat ctgtgtgttg gttttttgtg tgttcgaact agatgctgtc gaccgatgcc
     4801 cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc
     4861 cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt
     4921 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4981 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5041 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5101 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5161 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5221 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5281 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5341 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5401 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5461 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5521 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5581 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5641 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5701 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5761 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5821 caatctaaag tatatatgag taaacttggt ctgacagcgg ccgcaaatgc taaaccactg
     5881 cagtggttac cagtgcttga tcagtgaggc accgatctca gcgatctgcc tatttcgttc
     5941 gtccatagtg gcctgactcc ccgtcgtgta gatcactacg attcgtgagg gcttaccatc
     6001 aggccccagc gcagcaatga tgccgcgaga gccgcgttca ccggcccccg atttgtcagc
     6061 aatgaaccag ccagcaggga gggccgagcg aagaagtggt cctgctactt tgtccgcctc
     6121 catccagtct atgagctgct gtcgtgatgc tagagtaaga agttcgccag tgagtagttt
     6181 ccgaagagtt gtggccattg ctactggcat cgtggtatca cgctcgtcgt tcggtatggc
     6241 ttcgttcaac tc
//
