LOCUS       sequence-341625-        5553 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 5553)
  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 5553)
  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_S3
FEATURES             Location/Qualifiers
     source          1..5553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        585..781
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      632..767
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      811..820
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             817..2469
                     /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(883..901)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2510..2631
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2521..2540)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2575..2594
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2885..2902)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3056..3644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3136..3155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3815..4606,4607..4675))
                     /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     4438..4457
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4676..4780)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4807..5262
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4894..4913)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5104..5125
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5393..5441
                     /note="synthetic polyadenylation signal"
     misc_feature    5455..5546
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5495..5514
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtacctcag agagtagaaa ttgagggact taaaacattt tcttacgaga acatagaaaa
       61 aaaaacatat ctgaggcacg tgttttatga gattaagaag aattgctaaa caattgtaac
      121 ctaataccaa gacccagagg ggatggctca aggaaagaaa gttttcttga ttacagccta
      181 gttaagcaat ttaattaaaa tggctgcttt aaaaatatgt gcaattttgc caatggtaag
      241 tgtgttttct ggtgacatgt agaaacactg atgagtccca gccaggatac cattacaaag
      301 acaaaaggtc actgggctct ccttacaaca aacacatgtt ttccattgac tgtctccccc
      361 atggtgaatt tccttgccac tcattagaat ttattgatgc ataattcccc tgcaaaatag
      421 attatggaaa atgtagtctt acataatctt tgtcttcttt ctctgaagaa ctgagtgaaa
      481 ggtgtatcac agtggccatt tttgaggtga cgatgaggtt gaggaaagag gaaaggcggt
      541 gaggagctct tacgcgtgct agcccgggct cgagatctgc gatctgcatc tcaattagtc
      601 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
      661 ccattctccg ccccatcgct gactaatttt ttttatttat gcagaggccg aggccgcctc
      721 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
      781 aaagcttggc attccggtac tgttggtaaa gccaccatgg aagacgccaa aaacataaag
      841 aaaggcccgg cgccattcta tccgctggaa gatggaaccg ctggagagca actgcataag
      901 gctatgaaga gatacgccct ggttcctgga acaattgctt ttacagatgc acatatcgag
      961 gtggacatca cttacgctga gtacttcgaa atgtccgttc ggttggcaga agctatgaaa
     1021 cgatatgggc tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc
     1081 tttatgccgg tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc gaacgacatt
     1141 tataatgaac gtgaattgct caacagtatg ggcatttcgc agcctaccgt ggtgttcgtt
     1201 tccaaaaagg ggttgcaaaa aattttgaac gtgcaaaaaa agctcccaat catccaaaaa
     1261 attattatca tggattctaa aacggattac cagggatttc agtcgatgta cacgttcgtc
     1321 acatctcatc tacctcccgg ttttaatgaa tacgattttg tgccagagtc cttcgatagg
     1381 gacaagacaa ttgcactgat catgaactcc tctggatcta ctggtctgcc taaaggtgtc
     1441 gctctgcctc atagaactgc ctgcgtgaga ttctcgcatg ccagagatcc tatttttggc
     1501 aatcaaatca ttccggatac tgcgatttta agtgttgttc cattccatca cggttttgga
     1561 atgtttacta cactcggata tttgatatgt ggatttcgag tcgtcttaat gtatagattt
     1621 gaagaagagc tgtttctgag gagccttcag gattacaaga ttcaaagtgc gctgctggtg
     1681 ccaaccctat tctccttctt cgccaaaagc actctgattg acaaatacga tttatctaat
     1741 ttacacgaaa ttgcttctgg tggcgctccc ctctctaagg aagtcgggga agcggttgcc
     1801 aagaggttcc atctgccagg tatcaggcaa ggatatgggc tcactgagac tacatcagct
     1861 attctgatta cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt
     1921 tttgaagcga aggttgtgga tctggatacc gggaaaacgc tgggcgttaa tcaaagaggc
     1981 gaactgtgtg tgagaggtcc tatgattatg tccggttatg taaacaatcc ggaagcgacc
     2041 aacgccttga ttgacaagga tggatggcta cattctggag acatagctta ctgggacgaa
     2101 gacgaacact tcttcatcgt tgaccgcctg aagtctctga ttaagtacaa aggctatcag
     2161 gtggctcccg ctgaattgga atccatcttg ctccaacacc ccaacatctt cgacgcaggt
     2221 gtcgcaggtc ttcccgacga tgacgccggt gaacttcccg ccgccgttgt tgttttggag
     2281 cacggaaaga cgatgacgga aaaagagatc gtggattacg tcgccagtca agtaacaacc
     2341 gcgaaaaagt tgcgcggagg agttgtgttt gtggacgaag taccgaaagg tcttaccgga
     2401 aaactcgacg caagaaaaat cagagagatc ctcataaagg ccaagaaggg cggaaagatc
     2461 gccgtgtaat tctagagtcg gggcggccgg ccgcttcgag cagacatgat aagatacatt
     2521 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     2581 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2641 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     2701 taaaacctct acaaatgtgg taaaatcgat aaggatctgg atccgtcgac cgatgccctt
     2761 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
     2821 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctcttccg
     2881 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2941 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3001 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3061 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3121 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3181 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3241 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3301 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3361 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3421 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3481 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3541 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3601 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3661 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3721 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3781 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3841 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3901 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3961 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4021 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4081 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4141 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4201 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4261 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4321 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4381 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4441 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4501 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4561 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4621 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4681 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4741 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4801 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4861 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4921 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4981 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     5041 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5101 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     5161 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     5221 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttgccattcg ccattcaggc
     5281 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagcccaagc
     5341 taccatgata agtaagtaat attaaggtac gggaggtact tggagcggcc gcaataaaat
     5401 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag tactaacata
     5461 cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg tccccagtgc
     5521 aagtgcaggt gccagaacat ttctctatcg ata
//
