LOCUS       sequence-254873-        6036 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pGL3 basic luciferase reporter with co-expression of renilla 
            luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6036)
  TITLE     Emory Custom Cloning Core Plasmids - Oskar Laur
REFERENCE   2  (bases 1 to 6036)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGL3basic luciferase and  renilla_polyA
FEATURES             Location/Qualifiers
     source          1..6036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      82..91
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             88..1740
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(154..172)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1781..1902
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2150..2167)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2321..2909)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2401..2420)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3080..3871,3872..3940))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3703..3722
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3941..4045)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4072..4527
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4159..4178)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4369..4390
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4759..4880
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4896..5831)
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    5876..5924
                     /note="synthetic polyadenylation signal"
     misc_feature    5938..6029
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5978..5997
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc tgcgatctaa gtaagcttgg
       61 cattccggta ctgttggtaa agccaccatg gaagacgcca aaaacataaa gaaaggcccg
      121 gcgccattct atccgctgga agatggaacc gctggagagc aactgcataa ggctatgaag
      181 agatacgccc tggttcctgg aacaattgct tttacagatg cacatatcga ggtggacatc
      241 acttacgctg agtacttcga aatgtccgtt cggttggcag aagctatgaa acgatatggg
      301 ctgaatacaa atcacagaat cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg
      361 gtgttgggcg cgttatttat cggagttgca gttgcgcccg cgaacgacat ttataatgaa
      421 cgtgaattgc tcaacagtat gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag
      481 gggttgcaaa aaattttgaa cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc
      541 atggattcta aaacggatta ccagggattt cagtcgatgt acacgttcgt cacatctcat
      601 ctacctcccg gttttaatga atacgatttt gtgccagagt ccttcgatag ggacaagaca
      661 attgcactga tcatgaactc ctctggatct actggtctgc ctaaaggtgt cgctctgcct
      721 catagaactg cctgcgtgag attctcgcat gccagagatc ctatttttgg caatcaaatc
      781 attccggata ctgcgatttt aagtgttgtt ccattccatc acggttttgg aatgtttact
      841 acactcggat atttgatatg tggatttcga gtcgtcttaa tgtatagatt tgaagaagag
      901 ctgtttctga ggagccttca ggattacaag attcaaagtg cgctgctggt gccaacccta
      961 ttctccttct tcgccaaaag cactctgatt gacaaatacg atttatctaa tttacacgaa
     1021 attgcttctg gtggcgctcc cctctctaag gaagtcgggg aagcggttgc caagaggttc
     1081 catctgccag gtatcaggca aggatatggg ctcactgaga ctacatcagc tattctgatt
     1141 acacccgagg gggatgataa accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg
     1201 aaggttgtgg atctggatac cgggaaaacg ctgggcgtta atcaaagagg cgaactgtgt
     1261 gtgagaggtc ctatgattat gtccggttat gtaaacaatc cggaagcgac caacgccttg
     1321 attgacaagg atggatggct acattctgga gacatagctt actgggacga agacgaacac
     1381 ttcttcatcg ttgaccgcct gaagtctctg attaagtaca aaggctatca ggtggctccc
     1441 gctgaattgg aatccatctt gctccaacac cccaacatct tcgacgcagg tgtcgcaggt
     1501 cttcccgacg atgacgccgg tgaacttccc gccgccgttg ttgttttgga gcacggaaag
     1561 acgatgacgg aaaaagagat cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag
     1621 ttgcgcggag gagttgtgtt tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac
     1681 gcaagaaaaa tcagagagat cctcataaag gccaagaagg gcggaaagat cgccgtgtaa
     1741 ttctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt
     1801 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     1861 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     1921 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     1981 tacaaatgtg gtaaaatcga taaggatccg tcgaccgatg cccttgagag ccttcaaccc
     2041 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
     2101 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg
     2161 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2221 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2281 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2341 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2401 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2461 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2521 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2581 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2641 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2701 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2761 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2821 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2881 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2941 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3001 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3061 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3121 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3181 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     3241 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     3301 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     3361 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     3421 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     3481 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     3541 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     3601 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     3661 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     3721 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     3781 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     3841 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     3901 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     3961 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4021 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct
     4081 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4141 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4201 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4261 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4321 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4381 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4441 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4501 ttaacaaaat attaacgctt acaatttgcc attcgccatt caggctgcgc aactgttggg
     4561 aagggcgatc ggtgcgggcc tcttcgctat tacgccagcc caagctacca tgataagtaa
     4621 gtaatattaa ggtacgggag gtacttggag cggccgctta tcgattttac cacatttgta
     4681 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     4741 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4801 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4861 aaactcatca atgtatctta tcatgtctgc tcgaattatt gttcattttt gagaactcgc
     4921 tcaacgaacg atttgatata ttttcccatt tcatcaggtg catcttcttg cgaaaaatga
     4981 agacctttta ctttgacaaa ttcagtatta ggaaacttct tggcaccttc aacaatagca
     5041 ttggaaaaga atcctgggtc cgattcaata aacatttttg gtaaatcatc acttgcacgt
     5101 agataagcat tataattcct aacaatttgt acaacgtcag gtttaccacc ttttactaac
     5161 gggatttcac gaggccatga taatgttgga cgacgaactt cacctttctc tttgaatggt
     5221 tcaagatatg ctgcaaattc ttctggttct aactttctca tgatttttga tggcaacatg
     5281 gtttccacga agaagttatt ctccaaaacc attttttctc cttcttcaga tttgatcaac
     5341 gcaatatctt cttcaatatc aggccattca tcccatgatt caatcacatc tactacactt
     5401 tcagcgtgaa ctattgcttt gatcttatct tgatgctcat agctataatg aaatgccaaa
     5461 caagcacccc aatcatggcc gacaaaaatg atcttctttg gtaaattaag aagttcaaac
     5521 catgcagtaa gatatttgta atgatcaagt aacctataag aaccattacc agatttgcct
     5581 gatttgccca taccaataag gtctggtata atacaccgcg ctactggctc aatatgtggc
     5641 acaacatgtc gccataaata agaagaggcc gcgttaccat gtaaaaaaat aacagcattt
     5701 tctgcatgtt tttctgaatc ataataatta ataaatgaat caagaacatt catttgttta
     5761 catctggccc accactgcgg accagttatc atccgtttcc tttgttctgg atcataaact
     5821 ttcgaagtca tggtggcatg ctcgtggcgt ttaaacagac gcagcgtgcg gccgcaataa
     5881 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagtactaac
     5941 atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag
     6001 tgcaagtgca ggtgccagaa catttctcta tcgata
//
