LOCUS       sequence-341763-        5833 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Firefly luciferase enhancer reporter plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5833)
  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 5833)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_S5
FEATURES             Location/Qualifiers
     source          1..5833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        865..1061
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      912..1047
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      1091..1100
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1097..2749
                     /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(1163..1181)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2790..2911
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2801..2820)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2855..2874
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3165..3182)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3336..3924)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3416..3435)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4095..4886,4887..4955))
                     /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     4718..4737
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4956..5060)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5087..5542
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5174..5193)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5384..5405
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5673..5721
                     /note="synthetic polyadenylation signal"
     misc_feature    5735..5826
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5775..5794
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcctct acacagctgg tcaatccata gttgtttttt
       61 caattccagg ttcaagttac attttattgt ttattgtttt attttgaaat atttcttctt
      121 ggcaccctta ccagaaaagt cgcaatcttt ggtcgaaatt tgataatgga agattagttt
      181 actggtcagg atattgtgga ttaattaaga tggaaatctg gaaactggtt ttcattcttt
      241 gtttcctaca gtgagcagta aaagaacaaa ttaatggagt gaaacatgat ctcttggctg
      301 ctaaattcca aagaacaaat gtgatattta attacttaca gatattgagc ctttttcccc
      361 cactaatttt aattttgtct ccccatttgt aaattatttg gtgtttttgt ttgtttgtgc
      421 cattttgttt ttaagttaga actcatacaa aggtaatata tcaccataag ccctgagggc
      481 atgacacaga agattcactt acaaaggaca gacactcttt ctaatgggtc acaacattcc
      541 tctgatggat aggagtcact aaactgacaa taggttgctt gaaagagctc tcatctgagc
      601 tttaatcctt atttgattta acattttatc ttcattgaaa tatcttaaac atttgttgaa
      661 tgataaaatc aggatgtttt tatttgtatt ccctgagagc taaaattaaa cagaagatgc
      721 acgtatttca cttcccatat tttagagaaa ttatttgagc tacactaaat acagcattaa
      781 taaatctgac tgtggcttaa ctatatttgt tcttcttaat taactccaca aaatatctgc
      841 cacccgggct cgagatctgc gatctgcatc tcaattagtc agcaaccata gtcccgcccc
      901 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatcgct
      961 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     1021 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttggc attccggtac
     1081 tgttggtaaa gccaccatgg aagacgccaa aaacataaag aaaggcccgg cgccattcta
     1141 tccgctggaa gatggaaccg ctggagagca actgcataag gctatgaaga gatacgccct
     1201 ggttcctgga acaattgctt ttacagatgc acatatcgag gtggacatca cttacgctga
     1261 gtacttcgaa atgtccgttc ggttggcaga agctatgaaa cgatatgggc tgaatacaaa
     1321 tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc
     1381 gttatttatc ggagttgcag ttgcgcccgc gaacgacatt tataatgaac gtgaattgct
     1441 caacagtatg ggcatttcgc agcctaccgt ggtgttcgtt tccaaaaagg ggttgcaaaa
     1501 aattttgaac gtgcaaaaaa agctcccaat catccaaaaa attattatca tggattctaa
     1561 aacggattac cagggatttc agtcgatgta cacgttcgtc acatctcatc tacctcccgg
     1621 ttttaatgaa tacgattttg tgccagagtc cttcgatagg gacaagacaa ttgcactgat
     1681 catgaactcc tctggatcta ctggtctgcc taaaggtgtc gctctgcctc atagaactgc
     1741 ctgcgtgaga ttctcgcatg ccagagatcc tatttttggc aatcaaatca ttccggatac
     1801 tgcgatttta agtgttgttc cattccatca cggttttgga atgtttacta cactcggata
     1861 tttgatatgt ggatttcgag tcgtcttaat gtatagattt gaagaagagc tgtttctgag
     1921 gagccttcag gattacaaga ttcaaagtgc gctgctggtg ccaaccctat tctccttctt
     1981 cgccaaaagc actctgattg acaaatacga tttatctaat ttacacgaaa ttgcttctgg
     2041 tggcgctccc ctctctaagg aagtcgggga agcggttgcc aagaggttcc atctgccagg
     2101 tatcaggcaa ggatatgggc tcactgagac tacatcagct attctgatta cacccgaggg
     2161 ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga aggttgtgga
     2221 tctggatacc gggaaaacgc tgggcgttaa tcaaagaggc gaactgtgtg tgagaggtcc
     2281 tatgattatg tccggttatg taaacaatcc ggaagcgacc aacgccttga ttgacaagga
     2341 tggatggcta cattctggag acatagctta ctgggacgaa gacgaacact tcttcatcgt
     2401 tgaccgcctg aagtctctga ttaagtacaa aggctatcag gtggctcccg ctgaattgga
     2461 atccatcttg ctccaacacc ccaacatctt cgacgcaggt gtcgcaggtc ttcccgacga
     2521 tgacgccggt gaacttcccg ccgccgttgt tgttttggag cacggaaaga cgatgacgga
     2581 aaaagagatc gtggattacg tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg
     2641 agttgtgttt gtggacgaag taccgaaagg tcttaccgga aaactcgacg caagaaaaat
     2701 cagagagatc ctcataaagg ccaagaaggg cggaaagatc gccgtgtaat tctagagtcg
     2761 gggcggccgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     2821 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     2881 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     2941 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     3001 taaaatcgat aaggatctgg atccgtcgac cgatgccctt gagagccttc aacccagtca
     3061 gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta
     3121 tcatgcaact cgtaggacag gtgccggcag cgctcttccg cttcctcgct cactgactcg
     3181 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3241 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3301 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3361 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3421 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3481 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3541 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3601 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3661 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3721 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3781 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3841 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3901 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3961 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4021 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4081 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4141 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4201 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4261 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4321 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4381 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4441 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4501 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4561 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4621 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4681 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4741 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4801 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4861 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4921 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4981 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5041 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc
     5101 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     5161 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     5221 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     5281 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     5341 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5401 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     5461 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     5521 aaaatattaa cgcttacaat ttgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     5581 cgatcggtgc gggcctcttc gctattacgc cagcccaagc taccatgata agtaagtaat
     5641 attaaggtac gggaggtact tggagcggcc gcaataaaat atctttattt tcattacatc
     5701 tgtgtgttgg ttttttgtgt gaatcgatag tactaacata cgctctccat caaaacaaaa
     5761 cgaaacaaaa caaactagca aaataggctg tccccagtgc aagtgcaggt gccagaacat
     5821 ttctctatcg ata
//
