LOCUS       sequence-341761-        8544 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase enhancer reporter plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8544)
  AUTHORS   Long HK, Osterwalder M, Welsh IC, Hansen K, Davies JOJ, Liu YE, 
            Koska M, Adams AT, Aho R, Arora N, Ikeda K, Williams RM, 
            Sauka-Spengler T, Porteus MH, Mohun T, Dickel DE, Swigut T, Hughes 
            JR, Higgs DR, Visel A, Selleri L, Wysocka J
  TITLE     Loss of Extreme Long-Range Enhancers in Human Neural Crest Drives a 
            Craniofacial Disorder.
  JOURNAL   Cell Stem Cell. 2020 Nov 5;27(5):765-783.e14. doi: 
            10.1016/j.stem.2020.09.001. Epub 2020 Sep 28.
  PUBMED    32991838
REFERENCE   2  (bases 1 to 8544)
  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"; doi: 
            "10.1016/j.stem.2020.09.001"; journalName: "Cell Stem Cell"; date: 
            "2020-11-5- 5"; volume: "27"; issue: "5"; pages: "765-783.e14"
COMMENT     Sequence Label: pGL3-noSV40-humanEC1.45
FEATURES             Location/Qualifiers
     source          1..8544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        28..224
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      75..210
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      254..263
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             260..1912
                     /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(326..344)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1953..2074
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1964..1983)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2018..2037
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2328..2345)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2499..3087)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2579..2598)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3258..4049,4050..4118))
                     /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     3881..3900
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4119..4223)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4250..4705
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4337..4356)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4547..4568
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4836..4884
                     /note="synthetic polyadenylation signal"
     misc_feature    4898..4989
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4938..4957
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 gctagcccgg gctcgagatc tgcgatctgc atctcaatta gtcagcaacc atagtcccgc
       61 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatc
      121 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
      181 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt ggcattccgg
      241 tactgttggt aaagccacca tggaagacgc caaaaacata aagaaaggcc cggcgccatt
      301 ctatccgctg gaagatggaa ccgctggaga gcaactgcat aaggctatga agagatacgc
      361 cctggttcct ggaacaattg cttttacaga tgcacatatc gaggtggaca tcacttacgc
      421 tgagtacttc gaaatgtccg ttcggttggc agaagctatg aaacgatatg ggctgaatac
      481 aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg
      541 cgcgttattt atcggagttg cagttgcgcc cgcgaacgac atttataatg aacgtgaatt
      601 gctcaacagt atgggcattt cgcagcctac cgtggtgttc gtttccaaaa aggggttgca
      661 aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa aaaattatta tcatggattc
      721 taaaacggat taccagggat ttcagtcgat gtacacgttc gtcacatctc atctacctcc
      781 cggttttaat gaatacgatt ttgtgccaga gtccttcgat agggacaaga caattgcact
      841 gatcatgaac tcctctggat ctactggtct gcctaaaggt gtcgctctgc ctcatagaac
      901 tgcctgcgtg agattctcgc atgccagaga tcctattttt ggcaatcaaa tcattccgga
      961 tactgcgatt ttaagtgttg ttccattcca tcacggtttt ggaatgttta ctacactcgg
     1021 atatttgata tgtggatttc gagtcgtctt aatgtataga tttgaagaag agctgtttct
     1081 gaggagcctt caggattaca agattcaaag tgcgctgctg gtgccaaccc tattctcctt
     1141 cttcgccaaa agcactctga ttgacaaata cgatttatct aatttacacg aaattgcttc
     1201 tggtggcgct cccctctcta aggaagtcgg ggaagcggtt gccaagaggt tccatctgcc
     1261 aggtatcagg caaggatatg ggctcactga gactacatca gctattctga ttacacccga
     1321 gggggatgat aaaccgggcg cggtcggtaa agttgttcca ttttttgaag cgaaggttgt
     1381 ggatctggat accgggaaaa cgctgggcgt taatcaaaga ggcgaactgt gtgtgagagg
     1441 tcctatgatt atgtccggtt atgtaaacaa tccggaagcg accaacgcct tgattgacaa
     1501 ggatggatgg ctacattctg gagacatagc ttactgggac gaagacgaac acttcttcat
     1561 cgttgaccgc ctgaagtctc tgattaagta caaaggctat caggtggctc ccgctgaatt
     1621 ggaatccatc ttgctccaac accccaacat cttcgacgca ggtgtcgcag gtcttcccga
     1681 cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa agacgatgac
     1741 ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg
     1801 aggagttgtg tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa
     1861 aatcagagag atcctcataa aggccaagaa gggcggaaag atcgccgtgt aattctagag
     1921 tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     1981 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2041 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2101 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     2161 tggtaaaatc gataaggatc tggatccgtc gaccgatgcc cttgagagcc ttcaacccag
     2221 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     2281 ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac
     2341 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2401 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2461 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2521 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2581 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2641 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2701 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2761 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2821 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2881 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2941 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3001 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3061 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3121 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3181 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3241 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3301 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3361 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3421 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3481 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3541 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3601 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3661 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3721 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3781 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3841 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3901 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3961 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4021 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4081 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4141 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4201 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     4261 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4321 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4381 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4441 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4501 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4561 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4621 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4681 aacaaaatat taacgcttac aatttgccat tcgccattca ggctgcgcaa ctgttgggaa
     4741 gggcgatcgg tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt
     4801 aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta ttttcattac
     4861 atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca
     4921 aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa
     4981 catttctcta tcgataggta ccggacatcc tgattaagtg gcagcgagaa cttggaatat
     5041 ctgattaagt taaagatctt gagataggga ttatcctgaa tgatctggga ggggcccagt
     5101 gtgatcataa gaatccttat aaatcagaga gggagactgg aaggctagag tcagagagat
     5161 ttgcagatgc tatgttgtgg ctttgaagat ggaggaaggg gccatgagcc aaggaatgta
     5221 ttcagccccc agaaattaga aaagtcaggg aatagaatct gccctcgagc ctctggatgg
     5281 accgctcttc tgaggcctcc attttagccc attgcaaccc atttcagatt tctgacttga
     5341 gaccagtaag ataatacatt tgtgttgttt taagcagctg aatttgaggt aattagttac
     5401 agctgcaata ggtaaattga cattaaataa tatacttgac gttaggcttc tttactgtaa
     5461 ctcacttctg gatggcatgt tgttgtctgt ttttaccttt ctccttcccc cccccccagt
     5521 tgatctatta ggaaatagag aactatcttc atttctgctc cttcttccag gcttgaatct
     5581 acacaacagg ccactcccca aaattcctct gcttaattta gagagcaggt ttcagggatt
     5641 cacctgggaa caacagcagc aggcacctga ctcatattcc tggcggagag gaagggaggc
     5701 catgtagctg ctcttccgtg gacgcggatg cttcctgcct catagccggg tcgcctccgc
     5761 gctaaccccg ttgaaccttg caaccagccc ctctagagtg tggaagaaag gagcatcgac
     5821 ccagcctcag gtttcacttc tgttctagtt tatctatcta acttagacca catgtctgtg
     5881 ctttcagaaa attttcccat ttttgaatca ctaacagtac cctttattct tactactgcc
     5941 agggattcac tatttttaaa cagctttatt gtcatttcag taggatttgg ggaagaaaag
     6001 gaagcgaata catgtgttca atccgtcatg ttgacttgaa actgctgttt taaactgctc
     6061 tctttctacg tatttgattc cttttgctct acccatcccc atgtgcctct acttgtcaag
     6121 tattgggaga ttatataggt ttcttgtgtt tgttgtggtg atcaaagttt cttggggcag
     6181 cgccctcatt atctgtcagg cctggcttct atgattgaga acaggtgcag ccaagccttt
     6241 gctctcccta ccacttctgt ctgtcaaata gaaagcagga gggagctttt ctgtgggatc
     6301 tttcctttcc tctcaattct ctttgctgga gagctctcca ggatattggc aaggatctga
     6361 atttgtggct gaccaatggt tccacatcct gccatggccc ccacaaattt tgcagttttg
     6421 agatagtaga ttttgatgtg gttctgctgg agtcatatct cccatgtaat ttcaatttaa
     6481 tcagcttccg ttcatgcagt gaatgatggg ttcatttatc tgtgctttat ttgtatatga
     6541 tgaagtaatt ttttgaaaat tttatacata aatggtagca ttcccttatt tctaaatcag
     6601 atgctgggct tctctcaatc ttttgtatat tcctttgatt attagaatta taggtgggtg
     6661 aattcaagat aagagttaca catatgtgcc tgaattatca ctcacagacg atcagatatc
     6721 ttaatgtaat caataatctt atatccattc tctttttata agtaggtagg ttagaattca
     6781 gtttatccat tatgacttca ttttatagtt gaatcctcaa ttgcattaga agatatagag
     6841 gcctcactgt ctatttgcat gatgctttgg tgaattctgc tatattaggt tgttagggtc
     6901 actggggacc agaattattt gagaagactt agcatggagc atgtcatttc ttagaacatg
     6961 gctgtaaatg cttgttaaac gtgagaaaat ggggcatgtt aggaaagcca gaactggaaa
     7021 taaatccaaa aaattaaata tagctgattc tggggagata aatccatttt taatgataga
     7081 aggaaatcta atagacaatc taatagacaa aacatgttat ctaaagctca aggcaataga
     7141 caataagtgc tatatatatt tagggtccta actcaaaagt gtgccaaaag tgacagtggg
     7201 taaccaagag agagataaaa gatgtccgga ggtgttacag atacttggaa aagaagatac
     7261 acaagtagtt tcagaattct gtcagcaggg agagaaggct caggtttttt agctgaatag
     7321 caggaaagat cagtcccagg ggaagagaac tggctctgaa tgaaggaagt atatataggt
     7381 actaggcctt gtggatagga agagacaata agaagagcta gataattaca gaattttatc
     7441 tgtcacaata ttgcgtgctt ataacatcaa taaactaaaa gtaaaatcac aaaaccacca
     7501 actttacagt aagcttcttt gcaaattgta tggctttgtg tttaattaaa gcttactaac
     7561 gggttgggct cagatagttg tactcagaat acattttgac ttctttaagc ctaagataag
     7621 aatgttcatc ttcatttgga tggcatttat tacacaaacc caagaattcc atgatcatat
     7681 caataaaaat taggagaatt tcattaggca ctataaataa ccccttctcc tgagacaagt
     7741 agagaagccc gagtgagtaa ccaactgtcc ccaattgtat tagcctttta aataaaggat
     7801 tcccagcaaa cattcagaag aaaccatatg ccttccaggg ataggcagct ggaagagagg
     7861 gaagagatta atacagaaaa aagctatagg ccatcccgat cctattcttc agttccttta
     7921 ttggaggcat taagttttat tttattaact tctttcttgc ccttcaaaag ttgagggtgc
     7981 tcactccagg gcttgtggat atctgcccca gagaacctca gtgcaaaggt ttgaaatcac
     8041 agctggctgt gattagaagg attgggagag atcagtgctt gattaacttt cagctgctat
     8101 tggtttgcct cagttcctgc aggagaatta atctgtaggt tcgagtaaat tttggatctt
     8161 aatggatacc agatacattg agcccaccca gaccaccaag attttgcaat ccttttcctc
     8221 acatttaaat ggtacaaatg aattcattcc tccttactca gtgaacagaa taactctacc
     8281 tcaaggtttt tctcatatag cattcacttg attttgcctt cttttccaag aaagtataga
     8341 ttcgttgggg caaggtgcat tttcagaagt atgactttaa ttatttgcat tcagaggcat
     8401 ttgggtgaat aacaatgcat cctgctccct ttctcttaat ttgtgtttag tgcacataaa
     8461 attatgccag ctctgtaatc aataattttc attcttttgc acaagattca tcttcttcct
     8521 caactgaatt tatgccgtcc tagt
//
