LOCUS       sequence-341762-        5480 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 5480)
  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 5480)
  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_S4
FEATURES             Location/Qualifiers
     source          1..5480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        512..708
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      559..694
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      738..747
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             744..2396
                     /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(810..828)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2437..2558
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2448..2467)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2502..2521
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2812..2829)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2983..3571)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3063..3082)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3742..4533,4534..4602))
                     /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     4365..4384
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4603..4707)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4734..5189
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4821..4840)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5031..5052
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5320..5368
                     /note="synthetic polyadenylation signal"
     misc_feature    5382..5473
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5422..5441
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tctctcagga tgcaattgag ttacctacct ctgctgaaaa aatggattat
       61 tttgctaggt tgtgtgtggt gagttagact tatgacctag ggaagtagaa gtcaggaggg
      121 aaggcctggt agcagcatcc tataaaccag gccaatgtgt tgccctgaaa aggaatgcac
      181 tgccataaat gaaggcactt ggagggcaag ggtcatagta tgtaattagc gcaggccatt
      241 taaggaagaa taagaaaata atgttttaca agctggtaga aagagaagca aataatcatt
      301 taaatgttcc aattgtgagt tccgtcagtg tgtgtggttt attttataag caactttctg
      361 tactagtaag aggcaaagca atactgggaa atgaagtgct cattggtacc ctgcagtatg
      421 cactacaggg cccttccttt gaactgcttt gcataaggtt tatttccggc actctgtttt
      481 gttcgctagc ccgggctcga gatctgcgat ctgcatctca attagtcagc aaccatagtc
      541 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
      601 catcgctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
      661 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttggcatt
      721 ccggtactgt tggtaaagcc accatggaag acgccaaaaa cataaagaaa ggcccggcgc
      781 cattctatcc gctggaagat ggaaccgctg gagagcaact gcataaggct atgaagagat
      841 acgccctggt tcctggaaca attgctttta cagatgcaca tatcgaggtg gacatcactt
      901 acgctgagta cttcgaaatg tccgttcggt tggcagaagc tatgaaacga tatgggctga
      961 atacaaatca cagaatcgtc gtatgcagtg aaaactctct tcaattcttt atgccggtgt
     1021 tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg
     1081 aattgctcaa cagtatgggc atttcgcagc ctaccgtggt gttcgtttcc aaaaaggggt
     1141 tgcaaaaaat tttgaacgtg caaaaaaagc tcccaatcat ccaaaaaatt attatcatgg
     1201 attctaaaac ggattaccag ggatttcagt cgatgtacac gttcgtcaca tctcatctac
     1261 ctcccggttt taatgaatac gattttgtgc cagagtcctt cgatagggac aagacaattg
     1321 cactgatcat gaactcctct ggatctactg gtctgcctaa aggtgtcgct ctgcctcata
     1381 gaactgcctg cgtgagattc tcgcatgcca gagatcctat ttttggcaat caaatcattc
     1441 cggatactgc gattttaagt gttgttccat tccatcacgg ttttggaatg tttactacac
     1501 tcggatattt gatatgtgga tttcgagtcg tcttaatgta tagatttgaa gaagagctgt
     1561 ttctgaggag ccttcaggat tacaagattc aaagtgcgct gctggtgcca accctattct
     1621 ccttcttcgc caaaagcact ctgattgaca aatacgattt atctaattta cacgaaattg
     1681 cttctggtgg cgctcccctc tctaaggaag tcggggaagc ggttgccaag aggttccatc
     1741 tgccaggtat caggcaagga tatgggctca ctgagactac atcagctatt ctgattacac
     1801 ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg
     1861 ttgtggatct ggataccggg aaaacgctgg gcgttaatca aagaggcgaa ctgtgtgtga
     1921 gaggtcctat gattatgtcc ggttatgtaa acaatccgga agcgaccaac gccttgattg
     1981 acaaggatgg atggctacat tctggagaca tagcttactg ggacgaagac gaacacttct
     2041 tcatcgttga ccgcctgaag tctctgatta agtacaaagg ctatcaggtg gctcccgctg
     2101 aattggaatc catcttgctc caacacccca acatcttcga cgcaggtgtc gcaggtcttc
     2161 ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt tttggagcac ggaaagacga
     2221 tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc
     2281 gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa
     2341 gaaaaatcag agagatcctc ataaaggcca agaagggcgg aaagatcgcc gtgtaattct
     2401 agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat gagtttggac
     2461 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     2521 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     2581 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     2641 aatgtggtaa aatcgataag gatctggatc cgtcgaccga tgcccttgag agccttcaac
     2701 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc
     2761 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac
     2821 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2881 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2941 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3001 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3061 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3121 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3181 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3241 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3301 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3361 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3421 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3481 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3541 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3601 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3661 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3721 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3781 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3841 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3901 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3961 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4021 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4081 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4141 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4201 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4261 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4321 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4381 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4441 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4501 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4561 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4621 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4681 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc
     4741 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4801 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4861 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4921 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4981 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     5041 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     5101 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5161 ttttaacaaa atattaacgc ttacaatttg ccattcgcca ttcaggctgc gcaactgttg
     5221 ggaagggcga tcggtgcggg cctcttcgct attacgccag cccaagctac catgataagt
     5281 aagtaatatt aaggtacggg aggtacttgg agcggccgca ataaaatatc tttattttca
     5341 ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac taacatacgc tctccatcaa
     5401 aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc
     5461 agaacatttc tctatcgata
//
