LOCUS       sequence-341792-        5507 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 5507)
  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 5507)
  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_min1-2_4XCoordinatorMu...
FEATURES             Location/Qualifiers
     source          1..5507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1030..1151
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1041..1060)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1095..1114
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1405..1422)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1576..2164)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1656..1675)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2335..3126,3127..3195))
                     /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     2958..2977
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3196..3300)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3327..3782
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3414..3433)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3624..3645
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    3913..3961
                     /note="synthetic polyadenylation signal"
     misc_feature    3975..4066
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4015..4034
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        4612..4808
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4659..4794
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      4838..4847
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4844..5507,1..989)
                     /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(4910..4928)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
ORIGIN
        1 ccagagatcc tatttttggc aatcaaatca ttccggatac tgcgatttta agtgttgttc
       61 cattccatca cggttttgga atgtttacta cactcggata tttgatatgt ggatttcgag
      121 tcgtcttaat gtatagattt gaagaagagc tgtttctgag gagccttcag gattacaaga
      181 ttcaaagtgc gctgctggtg ccaaccctat tctccttctt cgccaaaagc actctgattg
      241 acaaatacga tttatctaat ttacacgaaa ttgcttctgg tggcgctccc ctctctaagg
      301 aagtcgggga agcggttgcc aagaggttcc atctgccagg tatcaggcaa ggatatgggc
      361 tcactgagac tacatcagct attctgatta cacccgaggg ggatgataaa ccgggcgcgg
      421 tcggtaaagt tgttccattt tttgaagcga aggttgtgga tctggatacc gggaaaacgc
      481 tgggcgttaa tcaaagaggc gaactgtgtg tgagaggtcc tatgattatg tccggttatg
      541 taaacaatcc ggaagcgacc aacgccttga ttgacaagga tggatggcta cattctggag
      601 acatagctta ctgggacgaa gacgaacact tcttcatcgt tgaccgcctg aagtctctga
      661 ttaagtacaa aggctatcag gtggctcccg ctgaattgga atccatcttg ctccaacacc
      721 ccaacatctt cgacgcaggt gtcgcaggtc ttcccgacga tgacgccggt gaacttcccg
      781 ccgccgttgt tgttttggag cacggaaaga cgatgacgga aaaagagatc gtggattacg
      841 tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt gtggacgaag
      901 taccgaaagg tcttaccgga aaactcgacg caagaaaaat cagagagatc ctcataaagg
      961 ccaagaaggg cggaaagatc gccgtgtaat tctagagtcg gggcggccgg ccgcttcgag
     1021 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     1081 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     1141 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     1201 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatctgg
     1261 atccgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     1321 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     1381 gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1441 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1501 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1561 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1621 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1681 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1741 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1801 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1861 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1921 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1981 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2041 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2101 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2161 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2221 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2281 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2341 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2401 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2461 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2521 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     2581 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2641 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     2701 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     2761 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     2821 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     2881 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     2941 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3001 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3061 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3121 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3181 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3241 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3301 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
     3361 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3421 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3481 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3541 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3601 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3661 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3721 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     3781 ttgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     3841 gctattacgc cagcccaagc taccatgata agtaagtaat attaaggtac gggaggtact
     3901 tggagcggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     3961 gaatcgatag tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca
     4021 aaataggctg tccccagtgc aagtgcaggt gccagaacat ttctctatcg ataggtacgc
     4081 catgtagctg ctcttccgtg gacgcggatg cttcctgcct catagccggg tcgcctccgc
     4141 gctaaccccg ttgaaccttg caaccagccc ctctagagtg tggaagaaag gagcatcgac
     4201 ccagcctcag gtttcacttc tgttctagtt tatctatccg gcttagattg agcatctgtg
     4261 ctttcagaaa attttcccat ttttgaatca ctaacagtac cctttattct tactactgcc
     4321 agggattcac tatttttaaa cagctcgagc ctcagtgcaa aggtttgaaa tcatgctcaa
     4381 ctgtggccgg aaggattggg agagatcagt gcttgattaa cttttgctca atattggccg
     4441 gcctcagttc ctgcaggaga attaatctgt aggttcgagt aaattttgga tctcggcgga
     4501 tattgagcac attgagccca cccagaccac caagattttg caatcctttt cctcacattt
     4561 aaatggtaca aatgaattca ttcctcctta ctcagtgaaa gatctgcgat ctgcatctca
     4621 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4681 gttccgccca ttctccgccc catcgctgac taattttttt tatttatgca gaggccgagg
     4741 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4801 tttgcaaaaa gcttggcatt ccggtactgt tggtaaagcc accatggaag acgccaaaaa
     4861 cataaagaaa ggcccggcgc cattctatcc gctggaagat ggaaccgctg gagagcaact
     4921 gcataaggct atgaagagat acgccctggt tcctggaaca attgctttta cagatgcaca
     4981 tatcgaggtg gacatcactt acgctgagta cttcgaaatg tccgttcggt tggcagaagc
     5041 tatgaaacga tatgggctga atacaaatca cagaatcgtc gtatgcagtg aaaactctct
     5101 tcaattcttt atgccggtgt tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa
     5161 cgacatttat aatgaacgtg aattgctcaa cagtatgggc atttcgcagc ctaccgtggt
     5221 gttcgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaagc tcccaatcat
     5281 ccaaaaaatt attatcatgg attctaaaac ggattaccag ggatttcagt cgatgtacac
     5341 gttcgtcaca tctcatctac ctcccggttt taatgaatac gattttgtgc cagagtcctt
     5401 cgatagggac aagacaattg cactgatcat gaactcctct ggatctactg gtctgcctaa
     5461 aggtgtcgct ctgcctcata gaactgcctg cgtgagattc tcgcatg
//
