LOCUS       sequence-341790-        6669 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 6669)
  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 6669)
  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-humanEC1.25_S6
FEATURES             Location/Qualifiers
     source          1..6669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1701..1897
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1748..1883
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      1927..1936
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1933..3585
                     /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(1999..2017)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3626..3747
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3637..3656)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3691..3710
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4001..4018)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4172..4760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4252..4271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4931..5722,5723..5791))
                     /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     5554..5573
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5792..5896)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5923..6378
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6010..6029)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6220..6241
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6509..6557
                     /note="synthetic polyadenylation signal"
     misc_feature    6571..6662
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6611..6630
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gttttgtggt gaatttcaag acttaaagta
       61 gctaatgcaa aaggatgata atttgtatta attttggtat actataccaa cagttttgaa
      121 taatatgtat gtcagaaggc cttaagaaat agagcattat gtctgctgta attattctct
      181 cctcttttct gccacacata gccacatggc tcagtcatta tcttctgcta tttagtatat
      241 ttaataccaa aggaattcct gctagccatc tgcttttcta tgttaattct tgccctttgc
      301 agttttcctt acaagatata atcatctaga acagcaggtc cctacagaga acttgaacat
      361 ttgcttttaa tcaaaaggca tagtttatat taaactcagt aaacaaaatc aggggacttg
      421 tataaataac aatttgtgca tatcattttc atttttgctg acacttttga ctctggtatt
      481 gatctttctc caggttgtga aactcagagg gcaaagtgca aagttcaaaa gtggctattt
      541 gcctcaaggt ttattgaaaa taaaacatca acggaagtcc tctgaagtaa tttgaggaaa
      601 cttttttttt cttctaggaa ggatgtcatt attcagtttc taaggttaag tctgcttaga
      661 gaaaaataat agagtgaaaa taggcttgaa acattagaaa gctggcagaa aacatctata
      721 taattgtggg actgaatgca agtatgtaaa atttccaaaa acaaacattg cattctatat
      781 ggaacatctc tttatctgtt catcgtccaa atacagaaac tatagctcca tcttgccaaa
      841 tccatagtat gtggactgaa aaaaaaaaaa aagaaaaaag aaaaaacaga tgcaaaatga
      901 ataaggaaag agtacattta aaaaaataca ttttttaaac tggtttaata agaaaattaa
      961 ttttaaaaaa tcagtacaat tggattgtca tcaagcagct aaaaggcaca gctcattttt
     1021 tcaattgtcc tgttccaaat gattattttg tttcatgcaa gcacttgctg ggaacgtttc
     1081 catgctgaaa caagacaagt gcaggcagag ctgtttttaa tttttaaaga taactaaaaa
     1141 catacagaaa agccttccaa gcttgacttg aaattgttgt atctaaagat tctcaatgca
     1201 tttattagga agtgtccatc caacttaaga gaattttaac aggcgtccta aggtcagact
     1261 gcaaatcagg ccaggagtca agagcttcag taatcttttc ccagcgtcat gcagtagatc
     1321 ctggttattt tcaaccaaat tagacatcat tttggatgtt tattctccca attatctcct
     1381 taccccccca atccttctaa gtttctttat ggttttattt aaaaattctc ttagtttttt
     1441 attgcgagag taacttaagt gtggtctatt tgaaatcttc atcacatata aagggggaac
     1501 acaaaatcat ttacgtaata agactccagc taacaggcac cctccagcaa tctgaatgct
     1561 tatgaaatgg catgaaaata aatccagaaa attaaactaa gagctagaca aaatataaac
     1621 tactttgagc tgtgctttta caagacagat atgtctaggt atatgagctc ctagttttcc
     1681 tggactcgag atctgcgatc tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     1741 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atcgctgact
     1801 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     1861 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttggcattc cggtactgtt
     1921 ggtaaagcca ccatggaaga cgccaaaaac ataaagaaag gcccggcgcc attctatccg
     1981 ctggaagatg gaaccgctgg agagcaactg cataaggcta tgaagagata cgccctggtt
     2041 cctggaacaa ttgcttttac agatgcacat atcgaggtgg acatcactta cgctgagtac
     2101 ttcgaaatgt ccgttcggtt ggcagaagct atgaaacgat atgggctgaa tacaaatcac
     2161 agaatcgtcg tatgcagtga aaactctctt caattcttta tgccggtgtt gggcgcgtta
     2221 tttatcggag ttgcagttgc gcccgcgaac gacatttata atgaacgtga attgctcaac
     2281 agtatgggca tttcgcagcc taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt
     2341 ttgaacgtgc aaaaaaagct cccaatcatc caaaaaatta ttatcatgga ttctaaaacg
     2401 gattaccagg gatttcagtc gatgtacacg ttcgtcacat ctcatctacc tcccggtttt
     2461 aatgaatacg attttgtgcc agagtccttc gatagggaca agacaattgc actgatcatg
     2521 aactcctctg gatctactgg tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc
     2581 gtgagattct cgcatgccag agatcctatt tttggcaatc aaatcattcc ggatactgcg
     2641 attttaagtg ttgttccatt ccatcacggt tttggaatgt ttactacact cggatatttg
     2701 atatgtggat ttcgagtcgt cttaatgtat agatttgaag aagagctgtt tctgaggagc
     2761 cttcaggatt acaagattca aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc
     2821 aaaagcactc tgattgacaa atacgattta tctaatttac acgaaattgc ttctggtggc
     2881 gctcccctct ctaaggaagt cggggaagcg gttgccaaga ggttccatct gccaggtatc
     2941 aggcaaggat atgggctcac tgagactaca tcagctattc tgattacacc cgagggggat
     3001 gataaaccgg gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt tgtggatctg
     3061 gataccggga aaacgctggg cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg
     3121 attatgtccg gttatgtaaa caatccggaa gcgaccaacg ccttgattga caaggatgga
     3181 tggctacatt ctggagacat agcttactgg gacgaagacg aacacttctt catcgttgac
     3241 cgcctgaagt ctctgattaa gtacaaaggc tatcaggtgg ctcccgctga attggaatcc
     3301 atcttgctcc aacaccccaa catcttcgac gcaggtgtcg caggtcttcc cgacgatgac
     3361 gccggtgaac ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa
     3421 gagatcgtgg attacgtcgc cagtcaagta acaaccgcga aaaagttgcg cggaggagtt
     3481 gtgtttgtgg acgaagtacc gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga
     3541 gagatcctca taaaggccaa gaagggcgga aagatcgccg tgtaattcta gagtcggggc
     3601 ggccggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     3661 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     3721 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     3781 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
     3841 atcgataagg atctggatcc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc
     3901 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     3961 gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc
     4021 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4081 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4141 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4201 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4261 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4321 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4381 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4441 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4501 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4561 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4621 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4681 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4741 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4801 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4861 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4921 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4981 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5041 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5101 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5161 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5221 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5281 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5341 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5401 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5461 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5521 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5581 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5641 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5701 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5761 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5821 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5881 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
     5941 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     6001 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     6061 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     6121 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     6181 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     6241 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     6301 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     6361 tattaacgct tacaatttgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     6421 cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc atgataagta agtaatatta
     6481 aggtacggga ggtacttgga gcggccgcaa taaaatatct ttattttcat tacatctgtg
     6541 tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa
     6601 acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct
     6661 ctatcgata
//
