LOCUS       sequence-127328-        6297 bp    DNA     circular SYN 14-APR-2025
DEFINITION  luciferase reposter with 229 bp of the HSV TK promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6297)
  AUTHORS   Hawwari A, Burrows J, Vadas MA, Cockerill PN
  TITLE     The human IL-3 locus is regulated cooperatively by two 
            NFAT-dependent enhancers that have distinct tissue-specific 
            activities.
  JOURNAL   J Immunol. 2002 Aug 15;169(4):1876-86.
  PUBMED    12165512
REFERENCE   2  (bases 1 to 6297)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J 
            Immunol."; date: "2002-08-15"; volume: "169(4)"; pages: "1876-86"
COMMENT     Sequence Label: pTK229
FEATURES             Location/Qualifiers
     source          1..6297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        105..250
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      280..289
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             286..1938
                     /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(352..370)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          2023..2088
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2218..2238
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2660..2794
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3565..3583)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3651..3755
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3756..3824,3825..4616)
                     /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(3974..3993)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4787..5375
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5276..5295
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5529..5546
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5561..5701
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5627..5646)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5787..5809
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     polyA_signal    5940..6074
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    join(6168..6297,1..5)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 ggatccaagc tcagatctcg agctgcttca tccccgtggc ccgttgctcg cgtttgctgg
       61 cggtgtcccc ggaagaaata tatttgcatg tctttagttc tatgatgaca caaaccccgc
      121 ccagcgtctt gtcattggcg aattcgaaca cgcagatgca gtcggggcgg cgcggtccca
      181 ggtccacttc gcatattaag gtgacgcgtg tggcctcgaa caccgagcga ccctgcagcg
      241 acccgcttaa aagcttggca ttccggtact gttggtaaag ccaccatgga agacgccaaa
      301 aacataaaga aaggcccggc gccattctat ccgctggaag atggaaccgc tggagagcaa
      361 ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt tacagatgca
      421 catatcgagg tggacatcac ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa
      481 gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag tgaaaactct
      541 cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg
      601 aacgacattt ataatgaacg tgaattgctc aacagtatgg gcatttcgca gcctaccgtg
      661 gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc
      721 atccaaaaaa ttattatcat ggattctaaa acggattacc agggatttca gtcgatgtac
      781 acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt gccagagtcc
      841 ttcgataggg acaagacaat tgcactgatc atgaactcct ctggatctac tggtctgcct
      901 aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc cagagatcct
      961 atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc attccatcac
     1021 ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt cgtcttaatg
     1081 tatagatttg aagaagagct gtttctgagg agccttcagg attacaagat tcaaagtgcg
     1141 ctgctggtgc caaccctatt ctccttcttc gccaaaagca ctctgattga caaatacgat
     1201 ttatctaatt tacacgaaat tgcttctggt ggcgctcccc tctctaagga agtcggggaa
     1261 gcggttgcca agaggttcca tctgccaggt atcaggcaag gatatgggct cactgagact
     1321 acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt cggtaaagtt
     1381 gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct gggcgttaat
     1441 caaagaggcg aactgtgtgt gagaggtcct atgattatgt ccggttatgt aaacaatccg
     1501 gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga catagcttac
     1561 tgggacgaag acgaacactt cttcatcgtt gaccgcctga agtctctgat taagtacaaa
     1621 ggctatcagg tggctcccgc tgaattggaa tccatcttgc tccaacaccc caacatcttc
     1681 gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg aacttcccgc cgccgttgtt
     1741 gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt cgccagtcaa
     1801 gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt accgaaaggt
     1861 cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc caagaagggc
     1921 ggaaagatcg ccgtgtaatt ctagaggatc tttgtgaagg aaccttactt ctgtggtgtg
     1981 acataattgg acaaactacc tacagagatt taaagctcta aggtaaatat aaaattttta
     2041 agtgtataat gtgttaaact actgattcta attgtttgtg tattttagat tccaacctat
     2101 ggaactgatg aatgggagca gtggtggaat gcctttaatg aggaaaacct gttttgctca
     2161 gaagaaatgc catctagtga tgatgaggct actgctgact ctcaacattc tactcctcca
     2221 aaaaagaaga gaaaggtaga agaccccaag gactttcctt cagaattgct aagttttttg
     2281 agtcatgctg tgtttagtaa tagaactctt gcttgctttg ctatttacac cacaaaggaa
     2341 aaagctgcac tgctatacaa gaaaattatg gaaaaatatt ctgtaacctt tataagtagg
     2401 cataacagtt ataatcataa catactgttt tttcttactc cacacaggca tagagtgtct
     2461 gctattaata actatgctca aaaattgtgt acctttagct ttttaatttg taaaggggtt
     2521 aataaggaat atttgatgta tagtgccttg actagagatc ataatcagcc ataccacatt
     2581 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2641 aatgaatgca attgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     2701 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2761 caaactcatc aatgtatctt atcatgtctg gatcgatccc caggaagctc ctctgtgtcc
     2821 tcataaaccc taacctcctc tacttgagag gacattccaa tcataggctg cccatccacc
     2881 ctctgtgtcc tcctgttaat taggtcactt aacaaaaagg aaattgggta ggggtttttc
     2941 acagaccgct ttctaagggt aattttaaaa tatctgggaa gtcccttcca ctgctgtgtt
     3001 ccagaagtgt tggtaaacag cccacaaatg tcaacagcag aaacatacaa gctgtcagct
     3061 ttgcacaagg gcccaacacc ctgctcatca agaagcactg tggttgctgt gttagtaatg
     3121 tgcaaaacag gaggcacatt ttccccacct gtgtaggttc caaaatatct agtgttttca
     3181 tttttacttg gatcaggaac ccagcactcc actggataag cattatcctt atccaaaaca
     3241 gccttgtggt cagtgttcat ctgctgactg tcaactgtag cattttttgg ggttacagtt
     3301 tgagcaggat atttggtcct gtagtttgct aacacaccct gcagctccaa aggttcccca
     3361 ccaacagcaa aaaaatgaaa atttgaccct tgaatgggtt ttccagcacc attttcatga
     3421 gttttttgtg tccctgaatg caagtttaac atagcagtta ccccaataac ctcagtttta
     3481 acagtaacag cttcccacat caaaatattt ccacaggtta agtcctcatt taaattaggc
     3541 aaaggaattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     3601 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3661 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3721 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3781 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3841 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3901 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3961 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     4021 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4081 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4141 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4201 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4261 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
     4321 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4381 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4441 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4501 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4561 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4621 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4681 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4741 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4801 ttctgcgcgt aatatgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4861 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4921 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4981 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5041 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5101 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5161 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5221 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5281 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5341 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5401 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5461 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     5521 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     5581 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     5641 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     5701 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     5761 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     5821 atcaccgaaa cgcgcgaggc aggatcagcc atatcacatt tgtagaggtt ttacttgctt
     5881 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgtta
     5941 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6001 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6061 atcatgtctg gatcagccat atcacatttg tagaggtttt acttgcttta aaaaacctcc
     6121 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttaac ttgtttattg
     6181 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6241 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtct
//
