LOCUS       sequence-341709-        6219 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 6219)
  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 6219)
  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-humanEC1.35_S1
FEATURES             Location/Qualifiers
     source          1..6219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1251..1447
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1298..1433
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      1477..1486
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1483..3135
                     /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(1549..1567)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3176..3297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3187..3206)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3241..3260
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3551..3568)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3722..4310)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3802..3821)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4481..5272,5273..5341))
                     /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     5104..5123
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5342..5446)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5473..5928
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5560..5579)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5770..5791
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6059..6107
                     /note="synthetic polyadenylation signal"
     misc_feature    6121..6212
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6161..6180
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtacctggg atgagagaga aagggctgtt ttgtctctca aggaattgct aaaccagatg
       61 ttgaaaaggg aggagagatt tttaaatttt agttgaaaag cagtatccaa tgaattaaaa
      121 actgataccc aatcagtaac aatgcctcca gccacatagc tttgggcact tctgagtcca
      181 cctctggcca cttctttact cctccatgaa gtgacttttc caggtcactt cttgttgtat
      241 atttgtggag gatttctact agaagcgtgc agatcaccat ctgctccaca gatcaaaggc
      301 atccatctcc taagttcccc actgaaacaa gggttaatgt tggaaaattt aattcagtga
      361 atgccactca taaaaatgaa ttgcagattt taatgaacct taattttcaa aggaatttat
      421 tctatgatat ttagctaaaa ataaaaatac aataatataa tggtataatt agttttctta
      481 aattagtata ttttctgttt ggtgcctgat agagggaaaa atggcattgt tgtacttcga
      541 agacgttttt cttcattctg tcttagcagg caaacttttt tttttttaaa tagtatttac
      601 acataatgaa atggttataa aaaatctaag cttattttca acacgtaaag agatgtttct
      661 gagtgatgtt tgtgtgttgt aaatttaagc aacctttcct taactttgaa atttttcaaa
      721 ctttgtatct ttgaaagatt tatgttaaaa gctgaaaaca tcaaaattaa gaagccgaac
      781 ccccaaattt ctgattttta agatggaaag ccatctgaac ttttacagga ttccaacagg
      841 gggcagtgaa attccacaga tgaacttgat ggctgtgctg aaaaatagat gcccagaatg
      901 taaatctctg aagagaaata tgtcttaaca cagctacctc aggaaagggg gaattattgc
      961 acttaattaa agtgtattga caccatatgt ggccccacct ctcccactcc cctttttgga
     1021 ttcattaaga attaaaaaaa aaaatcagaa actagtaaag gcaatattcc tatagagcct
     1081 gtgacttaac ccttaatcat taatcctaca aaaattacac ttaaacaaaa agctagttgg
     1141 taaacagaaa cctttcaaat tagacaattt atgttcatta tctctataac tcaaaggaat
     1201 gtcattggag agctcttacg cgtgctagcc cgggctcgag atctgcgatc tgcatctcaa
     1261 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     1321 ttccgcccat tctccgcccc atcgctgact aatttttttt atttatgcag aggccgaggc
     1381 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     1441 ttgcaaaaag cttggcattc cggtactgtt ggtaaagcca ccatggaaga cgccaaaaac
     1501 ataaagaaag gcccggcgcc attctatccg ctggaagatg gaaccgctgg agagcaactg
     1561 cataaggcta tgaagagata cgccctggtt cctggaacaa ttgcttttac agatgcacat
     1621 atcgaggtgg acatcactta cgctgagtac ttcgaaatgt ccgttcggtt ggcagaagct
     1681 atgaaacgat atgggctgaa tacaaatcac agaatcgtcg tatgcagtga aaactctctt
     1741 caattcttta tgccggtgtt gggcgcgtta tttatcggag ttgcagttgc gcccgcgaac
     1801 gacatttata atgaacgtga attgctcaac agtatgggca tttcgcagcc taccgtggtg
     1861 ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc aaaaaaagct cccaatcatc
     1921 caaaaaatta ttatcatgga ttctaaaacg gattaccagg gatttcagtc gatgtacacg
     1981 ttcgtcacat ctcatctacc tcccggtttt aatgaatacg attttgtgcc agagtccttc
     2041 gatagggaca agacaattgc actgatcatg aactcctctg gatctactgg tctgcctaaa
     2101 ggtgtcgctc tgcctcatag aactgcctgc gtgagattct cgcatgccag agatcctatt
     2161 tttggcaatc aaatcattcc ggatactgcg attttaagtg ttgttccatt ccatcacggt
     2221 tttggaatgt ttactacact cggatatttg atatgtggat ttcgagtcgt cttaatgtat
     2281 agatttgaag aagagctgtt tctgaggagc cttcaggatt acaagattca aagtgcgctg
     2341 ctggtgccaa ccctattctc cttcttcgcc aaaagcactc tgattgacaa atacgattta
     2401 tctaatttac acgaaattgc ttctggtggc gctcccctct ctaaggaagt cggggaagcg
     2461 gttgccaaga ggttccatct gccaggtatc aggcaaggat atgggctcac tgagactaca
     2521 tcagctattc tgattacacc cgagggggat gataaaccgg gcgcggtcgg taaagttgtt
     2581 ccattttttg aagcgaaggt tgtggatctg gataccggga aaacgctggg cgttaatcaa
     2641 agaggcgaac tgtgtgtgag aggtcctatg attatgtccg gttatgtaaa caatccggaa
     2701 gcgaccaacg ccttgattga caaggatgga tggctacatt ctggagacat agcttactgg
     2761 gacgaagacg aacacttctt catcgttgac cgcctgaagt ctctgattaa gtacaaaggc
     2821 tatcaggtgg ctcccgctga attggaatcc atcttgctcc aacaccccaa catcttcgac
     2881 gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac ttcccgccgc cgttgttgtt
     2941 ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc cagtcaagta
     3001 acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg acgaagtacc gaaaggtctt
     3061 accggaaaac tcgacgcaag aaaaatcaga gagatcctca taaaggccaa gaagggcgga
     3121 aagatcgccg tgtaattcta gagtcggggc ggccggccgc ttcgagcaga catgataaga
     3181 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3241 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3301 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     3361 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atctggatcc gtcgaccgat
     3421 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     3481 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     3541 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3601 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3661 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3721 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3781 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3841 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3901 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3961 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4021 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4081 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4141 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4201 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4261 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4321 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4381 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4441 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4501 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4561 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4621 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4681 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4741 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4801 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4861 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4921 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4981 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5041 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5101 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5161 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5221 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5281 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5341 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5401 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5461 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     5521 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     5581 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5641 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     5701 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5761 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     5821 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     5881 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttgc cattcgccat
     5941 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     6001 ccaagctacc atgataagta agtaatatta aggtacggga ggtacttgga gcggccgcaa
     6061 taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact
     6121 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
     6181 cagtgcaagt gcaggtgcca gaacatttct ctatcgata
//
