LOCUS       sequence-131890-        6187 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  luciferase reporter with hGM-CSF -55 to +28 minimal promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6187)
  AUTHORS   Bert AG, Burrows J, Osborne CS, Cockerill PN
  TITLE     Generation of an improved luciferase reporter gene plasmid that 
            employs a novel mechanism for high-copy replication.
  JOURNAL   Plasmid. 2000 Sep;44(2):173-82.
  PUBMED    10964627
REFERENCE   2  (bases 1 to 6187)
  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"; journalName: "Plasmid."; 
            date: "2000-09"; volume: "44(2)"; pages: "173-82"
COMMENT     Sequence Label: pGM55
FEATURES             Location/Qualifiers
     source          1..6187
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      170..179
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             176..1828
                     /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(242..260)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          1913..1978
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2108..2128
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2550..2684
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3455..3473)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3541..3645
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3646..3714,3715..4506)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3864..3883)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4677..5265
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5166..5185
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5419..5436
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5451..5591
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5517..5536)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5677..5699
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     polyA_signal    5830..5964
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    join(6058..6187,1..5)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 ggatccaagc tcagatctcg agctcggtac ccgggtcgac aagcttcgcg aggtcaccat
       61 taatcatttc ctctgtgtat ttaagagctc ttttgccagt gagcccagta cacagagaga
      121 aaggctaaag ttctgtcgac aagcttggca ttccggtact gttggtaaag ccaccatgga
      181 agacgccaaa aacataaaga aaggcccggc gccattctat ccgctggaag atggaaccgc
      241 tggagagcaa ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt
      301 tacagatgca catatcgagg tggacatcac ttacgctgag tacttcgaaa tgtccgttcg
      361 gttggcagaa gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag
      421 tgaaaactct cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt
      481 tgcgcccgcg aacgacattt ataatgaacg tgaattgctc aacagtatgg gcatttcgca
      541 gcctaccgtg gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa
      601 gctcccaatc atccaaaaaa ttattatcat ggattctaaa acggattacc agggatttca
      661 gtcgatgtac acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt
      721 gccagagtcc ttcgataggg acaagacaat tgcactgatc atgaactcct ctggatctac
      781 tggtctgcct aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc
      841 cagagatcct atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc
      901 attccatcac ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt
      961 cgtcttaatg tatagatttg aagaagagct gtttctgagg agccttcagg attacaagat
     1021 tcaaagtgcg ctgctggtgc caaccctatt ctccttcttc gccaaaagca ctctgattga
     1081 caaatacgat ttatctaatt tacacgaaat tgcttctggt ggcgctcccc tctctaagga
     1141 agtcggggaa gcggttgcca agaggttcca tctgccaggt atcaggcaag gatatgggct
     1201 cactgagact acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt
     1261 cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct
     1321 gggcgttaat caaagaggcg aactgtgtgt gagaggtcct atgattatgt ccggttatgt
     1381 aaacaatccg gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga
     1441 catagcttac tgggacgaag acgaacactt cttcatcgtt gaccgcctga agtctctgat
     1501 taagtacaaa ggctatcagg tggctcccgc tgaattggaa tccatcttgc tccaacaccc
     1561 caacatcttc gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg aacttcccgc
     1621 cgccgttgtt gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt
     1681 cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt
     1741 accgaaaggt cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc
     1801 caagaagggc ggaaagatcg ccgtgtaatt ctagaggatc tttgtgaagg aaccttactt
     1861 ctgtggtgtg acataattgg acaaactacc tacagagatt taaagctcta aggtaaatat
     1921 aaaattttta agtgtataat gtgttaaact actgattcta attgtttgtg tattttagat
     1981 tccaacctat ggaactgatg aatgggagca gtggtggaat gcctttaatg aggaaaacct
     2041 gttttgctca gaagaaatgc catctagtga tgatgaggct actgctgact ctcaacattc
     2101 tactcctcca aaaaagaaga gaaaggtaga agaccccaag gactttcctt cagaattgct
     2161 aagttttttg agtcatgctg tgtttagtaa tagaactctt gcttgctttg ctatttacac
     2221 cacaaaggaa aaagctgcac tgctatacaa gaaaattatg gaaaaatatt ctgtaacctt
     2281 tataagtagg cataacagtt ataatcataa catactgttt tttcttactc cacacaggca
     2341 tagagtgtct gctattaata actatgctca aaaattgtgt acctttagct ttttaatttg
     2401 taaaggggtt aataaggaat atttgatgta tagtgccttg actagagatc ataatcagcc
     2461 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     2521 tgaaacataa aatgaatgca attgttgtta acttgtttat tgcagcttat aatggttaca
     2581 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2641 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcgatccc caggaagctc
     2701 ctctgtgtcc tcataaaccc taacctcctc tacttgagag gacattccaa tcataggctg
     2761 cccatccacc ctctgtgtcc tcctgttaat taggtcactt aacaaaaagg aaattgggta
     2821 ggggtttttc acagaccgct ttctaagggt aattttaaaa tatctgggaa gtcccttcca
     2881 ctgctgtgtt ccagaagtgt tggtaaacag cccacaaatg tcaacagcag aaacatacaa
     2941 gctgtcagct ttgcacaagg gcccaacacc ctgctcatca agaagcactg tggttgctgt
     3001 gttagtaatg tgcaaaacag gaggcacatt ttccccacct gtgtaggttc caaaatatct
     3061 agtgttttca tttttacttg gatcaggaac ccagcactcc actggataag cattatcctt
     3121 atccaaaaca gccttgtggt cagtgttcat ctgctgactg tcaactgtag cattttttgg
     3181 ggttacagtt tgagcaggat atttggtcct gtagtttgct aacacaccct gcagctccaa
     3241 aggttcccca ccaacagcaa aaaaatgaaa atttgaccct tgaatgggtt ttccagcacc
     3301 attttcatga gttttttgtg tccctgaatg caagtttaac atagcagtta ccccaataac
     3361 ctcagtttta acagtaacag cttcccacat caaaatattt ccacaggtta agtcctcatt
     3421 taaattaggc aaaggaattc ttgaagacga aagggcctcg tgatacgcct atttttatag
     3481 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     3541 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3601 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3661 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3721 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3781 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3841 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     3901 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3961 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4021 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4081 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4141 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     4201 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4261 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4321 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4381 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4441 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     4501 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     4561 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4621 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4681 gatccttttt ttctgcgcgt aatatgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4741 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4801 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     4861 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4921 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4981 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5041 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5101 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5161 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5221 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5281 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5341 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     5401 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     5461 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     5521 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     5581 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     5641 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     5701 tttcaccgtc atcaccgaaa cgcgcgaggc aggatcagcc atatcacatt tgtagaggtt
     5761 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     5821 attgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5881 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5941 aatgtatctt atcatgtctg gatcagccat atcacatttg tagaggtttt acttgcttta
     6001 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttaac
     6061 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     6121 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6181 catgtct
//
