LOCUS       sequence-131892-        7477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  luciferase reporter with GM-CSF promoter + 717 bp Bgl II GM-CSF 
            enhancer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7477)
  AUTHORS   Cockerill PN, Bert AG, Roberts D, Vadas MA
  TITLE     The human granulocyte-macrophage colony-stimulating factor gene is 
            autonomously regulated in vivo by an inducible tissue-specific 
            enhancer.
  JOURNAL   Proc Natl Acad Sci U S A. 1999 Dec 21;96(26):15097-102.
  PUBMED    10611344
REFERENCE   2  (bases 1 to 7477)
  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: "Proc Natl 
            Acad Sci U S A."; date: "1999-12-21"; volume: "96(26)"; pages: 
            "15097-102"
COMMENT     Sequence Label: pGME
FEATURES             Location/Qualifiers
     source          1..7477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1460..1469
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1466..3118
                     /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(1532..1550)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          3203..3268
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3398..3418
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3840..3974
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4745..4763)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4831..4935
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4936..5004,5005..5796)
                     /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(5154..5173)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5967..6555
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6456..6475
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6709..6726
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6741..6881
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6807..6826)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6967..6989
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     polyA_signal    7120..7254
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    join(7348..7477,1..5)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 ggatccaagc tcagatctca ggtccccaga gataacaaga accctcatca agggttggac
       61 aggtcaaaga aggatcagat cagccccaaa gcctgtgcgg gggtgccatg gtaccgaagg
      121 gactttgagg cccatttcct tccccttgtg ataatgtctc tcgtgataag gatcctggag
      181 tgactcaagc ccctgtttcc tacagagcct gtggggtgga cacgcatagg aaactccttc
      241 cagagggttt tctcctgtct ctgtaggaag gggggcccca gaggggtcat accacccatc
      301 gtggtccccg cctctgccct gccacagccc catcggagcc cctgagtcag catggctggc
      361 tatcggttga cactgtttaa aatacagtgc aatctaggaa tgcctgcctg tgtcatggcc
      421 tcagccctga tgtcatcttt ccatgagaaa gatgtgtggc tgcccacaaa cctcatcacc
      481 aaacccatgg atttctgtcc tgtttgtgct gacttgggca cctgcaggcc accactgccg
      541 aaatggattt cttattatga ctcttgcttt cctcctttcc ttccctccta ggcccttggc
      601 cactggcctg ttacttgccc atctgttatg tcccagccct taaacacagc tttgctggcc
      661 ctcagctctt cttcctgaat aacttccgtg gcctcagcta ccacacagac atgtaccgct
      721 gccagatctc gagctcggta cccgggtcga caagcttcgc gaggtcacca gcttgctgag
      781 agtggctgca gtctcgctgc tggatgtgca catggtggtc attccctctg ctcacagggg
      841 caggggtccc cccttactgg actgaggttg ccccctgctc caggtcctgg gtgggagccc
      901 atgtgaactg tcagtggggc aggtctgtga gagctcccct cacactcaag tctctcacag
      961 tggccagaga agaggaaggc tggagtcaga atgaggcacc agggcgggca tagcctgccc
     1021 aaaggcccct gggattacag gcaggatggg gagccctatc taagtgtctc ccacgcccca
     1081 ccccagccat tccaggccag gaagtccaaa ctgtgcccct cagagggagg gggcagcctc
     1141 aggcccattc agactgccca gggagggctg gagagccctc aggaaggcga gtgggtgggc
     1201 tgtcggttct tggaaaggtt cattaatgaa aacccccaag cctgaccacc tagggaaaag
     1261 gctcaccgtt cccatgtgtg gctgataagg gccaggagat tccacagttc aggtagttcc
     1321 cccgcctccc tggcattttg tggtcaccat taatcatttc ctctgtgtat ttaagagctc
     1381 ttttgccagt gagcccagta cacagagaga aaggctaaag ttctgtcgac aagcttggca
     1441 ttccggtact gttggtaaag ccaccatgga agacgccaaa aacataaaga aaggcccggc
     1501 gccattctat ccgctggaag atggaaccgc tggagagcaa ctgcataagg ctatgaagag
     1561 atacgccctg gttcctggaa caattgcttt tacagatgca catatcgagg tggacatcac
     1621 ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac gatatgggct
     1681 gaatacaaat cacagaatcg tcgtatgcag tgaaaactct cttcaattct ttatgccggt
     1741 gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg aacgacattt ataatgaacg
     1801 tgaattgctc aacagtatgg gcatttcgca gcctaccgtg gtgttcgttt ccaaaaaggg
     1861 gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc atccaaaaaa ttattatcat
     1921 ggattctaaa acggattacc agggatttca gtcgatgtac acgttcgtca catctcatct
     1981 acctcccggt tttaatgaat acgattttgt gccagagtcc ttcgataggg acaagacaat
     2041 tgcactgatc atgaactcct ctggatctac tggtctgcct aaaggtgtcg ctctgcctca
     2101 tagaactgcc tgcgtgagat tctcgcatgc cagagatcct atttttggca atcaaatcat
     2161 tccggatact gcgattttaa gtgttgttcc attccatcac ggttttggaa tgtttactac
     2221 actcggatat ttgatatgtg gatttcgagt cgtcttaatg tatagatttg aagaagagct
     2281 gtttctgagg agccttcagg attacaagat tcaaagtgcg ctgctggtgc caaccctatt
     2341 ctccttcttc gccaaaagca ctctgattga caaatacgat ttatctaatt tacacgaaat
     2401 tgcttctggt ggcgctcccc tctctaagga agtcggggaa gcggttgcca agaggttcca
     2461 tctgccaggt atcaggcaag gatatgggct cactgagact acatcagcta ttctgattac
     2521 acccgagggg gatgataaac cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa
     2581 ggttgtggat ctggataccg ggaaaacgct gggcgttaat caaagaggcg aactgtgtgt
     2641 gagaggtcct atgattatgt ccggttatgt aaacaatccg gaagcgacca acgccttgat
     2701 tgacaaggat ggatggctac attctggaga catagcttac tgggacgaag acgaacactt
     2761 cttcatcgtt gaccgcctga agtctctgat taagtacaaa ggctatcagg tggctcccgc
     2821 tgaattggaa tccatcttgc tccaacaccc caacatcttc gacgcaggtg tcgcaggtct
     2881 tcccgacgat gacgccggtg aacttcccgc cgccgttgtt gttttggagc acggaaagac
     2941 gatgacggaa aaagagatcg tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt
     3001 gcgcggagga gttgtgtttg tggacgaagt accgaaaggt cttaccggaa aactcgacgc
     3061 aagaaaaatc agagagatcc tcataaaggc caagaagggc ggaaagatcg ccgtgtaatt
     3121 ctagaggatc tttgtgaagg aaccttactt ctgtggtgtg acataattgg acaaactacc
     3181 tacagagatt taaagctcta aggtaaatat aaaattttta agtgtataat gtgttaaact
     3241 actgattcta attgtttgtg tattttagat tccaacctat ggaactgatg aatgggagca
     3301 gtggtggaat gcctttaatg aggaaaacct gttttgctca gaagaaatgc catctagtga
     3361 tgatgaggct actgctgact ctcaacattc tactcctcca aaaaagaaga gaaaggtaga
     3421 agaccccaag gactttcctt cagaattgct aagttttttg agtcatgctg tgtttagtaa
     3481 tagaactctt gcttgctttg ctatttacac cacaaaggaa aaagctgcac tgctatacaa
     3541 gaaaattatg gaaaaatatt ctgtaacctt tataagtagg cataacagtt ataatcataa
     3601 catactgttt tttcttactc cacacaggca tagagtgtct gctattaata actatgctca
     3661 aaaattgtgt acctttagct ttttaatttg taaaggggtt aataaggaat atttgatgta
     3721 tagtgccttg actagagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     3781 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgtta
     3841 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3901 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3961 atcatgtctg gatcgatccc caggaagctc ctctgtgtcc tcataaaccc taacctcctc
     4021 tacttgagag gacattccaa tcataggctg cccatccacc ctctgtgtcc tcctgttaat
     4081 taggtcactt aacaaaaagg aaattgggta ggggtttttc acagaccgct ttctaagggt
     4141 aattttaaaa tatctgggaa gtcccttcca ctgctgtgtt ccagaagtgt tggtaaacag
     4201 cccacaaatg tcaacagcag aaacatacaa gctgtcagct ttgcacaagg gcccaacacc
     4261 ctgctcatca agaagcactg tggttgctgt gttagtaatg tgcaaaacag gaggcacatt
     4321 ttccccacct gtgtaggttc caaaatatct agtgttttca tttttacttg gatcaggaac
     4381 ccagcactcc actggataag cattatcctt atccaaaaca gccttgtggt cagtgttcat
     4441 ctgctgactg tcaactgtag cattttttgg ggttacagtt tgagcaggat atttggtcct
     4501 gtagtttgct aacacaccct gcagctccaa aggttcccca ccaacagcaa aaaaatgaaa
     4561 atttgaccct tgaatgggtt ttccagcacc attttcatga gttttttgtg tccctgaatg
     4621 caagtttaac atagcagtta ccccaataac ctcagtttta acagtaacag cttcccacat
     4681 caaaatattt ccacaggtta agtcctcatt taaattaggc aaaggaattc ttgaagacga
     4741 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     4801 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4861 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4921 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4981 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5041 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5101 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5161 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     5221 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5281 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5341 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5401 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5461 cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5521 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5581 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5641 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5701 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5761 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5821 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5881 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5941 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatatgctgc
     6001 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6061 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     6121 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6181 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6241 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6301 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6361 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6421 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6481 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6541 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6601 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6661 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     6721 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     6781 tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     6841 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     6901 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     6961 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     7021 aggatcagcc atatcacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     7081 cccctgaacc tgaaacataa aatgaatgca attgttgtta acttgtttat tgcagcttat
     7141 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     7201 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcagccat
     7261 atcacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     7321 aaacataaaa tgaatgcaat tgttgttaac ttgtttattg cagcttataa tggttacaaa
     7381 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     7441 ggtttgtcca aactcatcaa tgtatcttat catgtct
//
