LOCUS       sequence-341650-        8500 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 8500)
  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 8500)
  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_S3-4-5-6
FEATURES             Location/Qualifiers
     source          1..8500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3532..3728
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3579..3714
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3758..3767
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3764..5416
                     /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(3830..3848)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    5457..5578
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5468..5487)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5522..5541
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5832..5849)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6003..6591)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6083..6102)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6762..7553,7554..7622))
                     /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     7385..7404
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7623..7727)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7754..8209
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7841..7860)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8051..8072
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    8340..8388
                     /note="synthetic polyadenylation signal"
     misc_feature    8402..8493
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     8442..8461
                     /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 ctcaggatgc aattgagtta cctacctctg ctgaaaaaat ggattatttt
      601 gctaggttgt gtgtggtgag ttagacttat gacctaggga agtagaagtc aggagggaag
      661 gcctggtagc agcatcctat aaaccaggcc aatgtgttgc cctgaaaagg aatgcactgc
      721 cataaatgaa ggcacttgga gggcaagggt catagtatgt aattagcgca ggccatttaa
      781 ggaagaataa gaaaataatg ttttacaagc tggtagaaag agaagcaaat aatcatttaa
      841 atgttccaat tgtgagttcc gtcagtgtgt gtggtttatt ttataagcaa ctttctgtac
      901 tagtaagagg caaagcaata ctgggaaatg aagtgctcat tggtaccctg cagtatgcac
      961 tacagggccc ttcctttgaa ctgctttgca taaggtttat ttccggcact ctgttttgtt
     1021 cgctagcccc gcgtgctagc ctctacacag ctggtcaatc catagttgtt ttttcaattc
     1081 caggttcaag ttacatttta ttgtttattg ttttattttg aaatatttct tcttggcacc
     1141 cttaccagaa aagtcgcaat ctttggtcga aatttgataa tggaagatta gtttactggt
     1201 caggatattg tggattaatt aagatggaaa tctggaaact ggttttcatt ctttgtttcc
     1261 tacagtgagc agtaaaagaa caaattaatg gagtgaaaca tgatctcttg gctgctaaat
     1321 tccaaagaac aaatgtgata tttaattact tacagatatt gagccttttt cccccactaa
     1381 ttttaatttt gtctccccat ttgtaaatta tttggtgttt ttgtttgttt gtgccatttt
     1441 gtttttaagt tagaactcat acaaaggtaa tatatcacca taagccctga gggcatgaca
     1501 cagaagattc acttacaaag gacagacact ctttctaatg ggtcacaaca ttcctctgat
     1561 ggataggagt cactaaactg acaataggtt gcttgaaaga gctctcatct gagctttaat
     1621 ccttatttga tttaacattt tatcttcatt gaaatatctt aaacatttgt tgaatgataa
     1681 aatcaggatg tttttatttg tattccctga gagctaaaat taaacagaag atgcacgtat
     1741 ttcacttccc atattttaga gaaattattt gagctacact aaatacagca ttaataaatc
     1801 tgactgtggc ttaactatat ttgttcttct taattaactc cacaaaatat ctgccacccg
     1861 ggttttgtgg tgaatttcaa gacttaaagt agctaatgca aaaggatgat aatttgtatt
     1921 aattttggta tactatacca acagttttga ataatatgta tgtcagaagg ccttaagaaa
     1981 tagagcatta tgtctgctgt aattattctc tcctcttttc tgccacacat agccacatgg
     2041 ctcagtcatt atcttctgct atttagtata tttaatacca aaggaattcc tgctagccat
     2101 ctgcttttct atgttaattc ttgccctttg cagttttcct tacaagatat aatcatctag
     2161 aacagcaggt ccctacagag aacttgaaca tttgctttta atcaaaaggc atagtttata
     2221 ttaaactcag taaacaaaat caggggactt gtataaataa caatttgtgc atatcatttt
     2281 catttttgct gacacttttg actctggtat tgatctttct ccaggttgtg aaactcagag
     2341 ggcaaagtgc aaagttcaaa agtggctatt tgcctcaagg tttattgaaa ataaaacatc
     2401 aacggaagtc ctctgaagta atttgaggaa actttttttt tcttctagga aggatgtcat
     2461 tattcagttt ctaaggttaa gtctgcttag agaaaaataa tagagtgaaa ataggcttga
     2521 aacattagaa agctggcaga aaacatctat ataattgtgg gactgaatgc aagtatgtaa
     2581 aatttccaaa aacaaacatt gcattctata tggaacatct ctttatctgt tcatcgtcca
     2641 aatacagaaa ctatagctcc atcttgccaa atccatagta tgtggactga aaaaaaaaaa
     2701 aaagaaaaaa gaaaaaacag atgcaaaatg aataaggaaa gagtacattt aaaaaaatac
     2761 attttttaaa ctggtttaat aagaaaatta attttaaaaa atcagtacaa ttggattgtc
     2821 atcaagcagc taaaaggcac agctcatttt ttcaattgtc ctgttccaaa tgattatttt
     2881 gtttcatgca agcacttgct gggaacgttt ccatgctgaa acaagacaag tgcaggcaga
     2941 gctgttttta atttttaaag ataactaaaa acatacagaa aagccttcca agcttgactt
     3001 gaaattgttg tatctaaaga ttctcaatgc atttattagg aagtgtccat ccaacttaag
     3061 agaattttaa caggcgtcct aaggtcagac tgcaaatcag gccaggagtc aagagcttca
     3121 gtaatctttt cccagcgtca tgcagtagat cctggttatt ttcaaccaaa ttagacatca
     3181 ttttggatgt ttattctccc aattatctcc ttaccccccc aatccttcta agtttcttta
     3241 tggttttatt taaaaattct cttagttttt tattgcgaga gtaacttaag tgtggtctat
     3301 ttgaaatctt catcacatat aaagggggaa cacaaaatca tttacgtaat aagactccag
     3361 ctaacaggca ccctccagca atctgaatgc ttatgaaatg gcatgaaaat aaatccagaa
     3421 aattaaacta agagctagac aaaatataaa ctactttgag ctgtgctttt acaagacaga
     3481 tatgtctagg tatatgagct cctagttttc ctggactcga gatctgcgat ctgcatctca
     3541 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3601 gttccgccca ttctccgccc catcgctgac taattttttt tatttatgca gaggccgagg
     3661 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3721 tttgcaaaaa gcttggcatt ccggtactgt tggtaaagcc accatggaag acgccaaaaa
     3781 cataaagaaa ggcccggcgc cattctatcc gctggaagat ggaaccgctg gagagcaact
     3841 gcataaggct atgaagagat acgccctggt tcctggaaca attgctttta cagatgcaca
     3901 tatcgaggtg gacatcactt acgctgagta cttcgaaatg tccgttcggt tggcagaagc
     3961 tatgaaacga tatgggctga atacaaatca cagaatcgtc gtatgcagtg aaaactctct
     4021 tcaattcttt atgccggtgt tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa
     4081 cgacatttat aatgaacgtg aattgctcaa cagtatgggc atttcgcagc ctaccgtggt
     4141 gttcgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaagc tcccaatcat
     4201 ccaaaaaatt attatcatgg attctaaaac ggattaccag ggatttcagt cgatgtacac
     4261 gttcgtcaca tctcatctac ctcccggttt taatgaatac gattttgtgc cagagtcctt
     4321 cgatagggac aagacaattg cactgatcat gaactcctct ggatctactg gtctgcctaa
     4381 aggtgtcgct ctgcctcata gaactgcctg cgtgagattc tcgcatgcca gagatcctat
     4441 ttttggcaat caaatcattc cggatactgc gattttaagt gttgttccat tccatcacgg
     4501 ttttggaatg tttactacac tcggatattt gatatgtgga tttcgagtcg tcttaatgta
     4561 tagatttgaa gaagagctgt ttctgaggag ccttcaggat tacaagattc aaagtgcgct
     4621 gctggtgcca accctattct ccttcttcgc caaaagcact ctgattgaca aatacgattt
     4681 atctaattta cacgaaattg cttctggtgg cgctcccctc tctaaggaag tcggggaagc
     4741 ggttgccaag aggttccatc tgccaggtat caggcaagga tatgggctca ctgagactac
     4801 atcagctatt ctgattacac ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt
     4861 tccatttttt gaagcgaagg ttgtggatct ggataccggg aaaacgctgg gcgttaatca
     4921 aagaggcgaa ctgtgtgtga gaggtcctat gattatgtcc ggttatgtaa acaatccgga
     4981 agcgaccaac gccttgattg acaaggatgg atggctacat tctggagaca tagcttactg
     5041 ggacgaagac gaacacttct tcatcgttga ccgcctgaag tctctgatta agtacaaagg
     5101 ctatcaggtg gctcccgctg aattggaatc catcttgctc caacacccca acatcttcga
     5161 cgcaggtgtc gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt
     5221 tttggagcac ggaaagacga tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt
     5281 aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct
     5341 taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca agaagggcgg
     5401 aaagatcgcc gtgtaattct agagtcgggg cggccggccg cttcgagcag acatgataag
     5461 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     5521 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     5581 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     5641 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatctggatc cgtcgaccga
     5701 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
     5761 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
     5821 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5881 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5941 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6001 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6061 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6121 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6181 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6241 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6301 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6361 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6421 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6481 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6541 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6601 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6661 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6721 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6781 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6841 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6901 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6961 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7021 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7081 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7141 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7201 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7261 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7321 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7381 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7441 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7501 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7561 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7621 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7681 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7741 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     7801 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     7861 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     7921 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     7981 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     8041 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     8101 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     8161 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg ccattcgcca
     8221 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     8281 cccaagctac catgataagt aagtaatatt aaggtacggg aggtacttgg agcggccgca
     8341 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac
     8401 taacatacgc tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc
     8461 ccagtgcaag tgcaggtgcc agaacatttc tctatcgata
//
