LOCUS       sequence-399715-        6307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pGL3 luciferase reporter with CHAF1B SVA cloned upstream of SV40 
            promoter to test effects of SVA insertion on luciferase expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6307)
  AUTHORS   Nadler MJS, Chang W, Ozkaynak E, Huo Y, Nong Y, Boillot M, Johnson 
            M, Moreno A, Matthew P Anderson
  TITLE     Hominoid SVA-lncRNA AK057321 targets human-specific SVA 
            retrotransposons in SCN8A and CDK5RAP2 to initiate neuronal 
            maturation.
  JOURNAL   Commun Biol. 2023 Mar 30;6(1):347. doi: 10.1038/s42003-023-04683-8.
  PUBMED    36997626
REFERENCE   2  (bases 1 to 6307)
  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"; journalName: "Commun 
            Biol."; date: "2023-03-30"; volume: "6(1):347. doi"; pages: " 
            10.1038/s42003-023-04683-8"
COMMENT     Sequence Label: pGL3-luciferase-reporter-CHAF1B-SVA
FEATURES             Location/Qualifiers
     source          1..6307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(73..92)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(752..1543,1544..1612))
                     /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     1375..1394
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1613..1717)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1744..2199
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1831..1850)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2041..2062
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2330..2378
                     /note="synthetic polyadenylation signal"
     misc_feature    2392..2483
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2432..2451
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        3835..4031
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3882..4017
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      4061..4070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4067..5719
                     /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(4133..4151)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    5760..5881
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5771..5790)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5825..5844
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6129..6146)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6300..6307,1..581))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
       61 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
      121 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
      181 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
      241 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
      301 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
      361 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
      421 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
      481 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
      541 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
      601 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
      661 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
      721 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
      781 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
      841 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
      901 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
      961 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     1021 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     1081 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     1141 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     1201 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     1261 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     1321 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     1381 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     1441 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     1501 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     1561 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     1621 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     1681 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     1741 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     1801 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     1861 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     1921 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     1981 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2041 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     2101 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     2161 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg ccattcgcca ttcaggctgc
     2221 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag cccaagctac
     2281 catgataagt aagtaatatt aaggtacggg aggtacttgg agcggccgca ataaaatatc
     2341 tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac taacatacgc
     2401 tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag
     2461 tgcaggtgcc agaacatttc tctatcgata ggtacccctc tccctctccc tccaccgtct
     2521 ccctctccct ctcccgctct ctccacagtc tccctctgat gccaagccga ggctggacgg
     2581 tactgccgcc atctcggctc actgcaacct ccctgcctga ttctcctgcc tcagcctgcc
     2641 gagtgcctgg gattgcaggc gcacgccgcc acgcctgtct ggttttcgta tttttttggt
     2701 ggagatgggg tttcgctgtg ttggccgggc tggtctccag ctcctaatcg cgagtgatct
     2761 gccagcctca gcctcccgag gtgccgggat tgcagacaga gtctcactca ctcagtgctc
     2821 aatgttgccc aggctggagg gcactggcgt ggtctccgct cgctacaacc tccacctccc
     2881 agccgcctgc cttggcctcc caaagtgctg agattgcagc ctctgcccgg ccgccacccc
     2941 gtctaggaag tgagtagcgt ctctgcccgg ccgcccatcg tctgagatgt ggggagcgcc
     3001 tctgccccgc cgccctgtct gggaggtgag gagcgtctct gcccggccgc cccgtctgag
     3061 aagtgaggag cccctccgcc cggcagccgc cccgtctggg aagagaggag cccctccgcc
     3121 cagcagccgc cccgtccggg aagagaggag catctctgcc cggcagctgc cccgtccagg
     3181 aggtgggggg cagcccccgc ccggccagct gccccgtccg ggaggtgggg ggcgcctctg
     3241 cccggccgcc ctgtctggga agtgaggagc ccctctaccc ggccgccacc ccgtctggga
     3301 ggtgtaccca acagctcact gagaacgggc catgatgacg atggcggttt tgtcgaatag
     3361 aaaaagggga aatgtgggga aaagatagag aaatcagatt gttgctgtgt ctgtgtagaa
     3421 agaggtagac ataggagact ccattttgtt ctgtactaag aaaaattctt ctgccttggg
     3481 atgctgctaa tctataacct tacccccaac cccctgctct ctgaaacatg tgctgtgtca
     3541 actcagggtt aaatggatta aggacggtac aagatgtgct ttgttaaaca gatgcttgaa
     3601 ggcagcatgc tccttcagag tcatcaccac tccctaatct cacgtaccca gggacacaaa
     3661 cactgcggaa ggccactggg tcctctgcct aggaaaacca gagacccttg ttcacttgtt
     3721 tatctgctga ccttccctcc actattgtcc tatgaccctg ccaaatctcc ctctccgaga
     3781 aacacccaag aatgatcaat aaatactgct agcccgggct cgagatctgc gatctgcatc
     3841 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3901 ccagttccgc ccattctccg ccccatcgct gactaatttt ttttatttat gcagaggccg
     3961 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4021 gcttttgcaa aaagcttggc attccggtac tgttggtaaa gccaccatgg aagacgccaa
     4081 aaacataaag aaaggcccgg cgccattcta tccgctggaa gatggaaccg ctggagagca
     4141 actgcataag gctatgaaga gatacgccct ggttcctgga acaattgctt ttacagatgc
     4201 acatatcgag gtggacatca cttacgctga gtacttcgaa atgtccgttc ggttggcaga
     4261 agctatgaaa cgatatgggc tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc
     4321 tcttcaattc tttatgccgg tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc
     4381 gaacgacatt tataatgaac gtgaattgct caacagtatg ggcatttcgc agcctaccgt
     4441 ggtgttcgtt tccaaaaagg ggttgcaaaa aattttgaac gtgcaaaaaa agctcccaat
     4501 catccaaaaa attattatca tggattctaa aacggattac cagggatttc agtcgatgta
     4561 cacgttcgtc acatctcatc tacctcccgg ttttaatgaa tacgattttg tgccagagtc
     4621 cttcgatagg gacaagacaa ttgcactgat catgaactcc tctggatcta ctggtctgcc
     4681 taaaggtgtc gctctgcctc atagaactgc ctgcgtgaga ttctcgcatg ccagagatcc
     4741 tatttttggc aatcaaatca ttccggatac tgcgatttta agtgttgttc cattccatca
     4801 cggttttgga atgtttacta cactcggata tttgatatgt ggatttcgag tcgtcttaat
     4861 gtatagattt gaagaagagc tgtttctgag gagccttcag gattacaaga ttcaaagtgc
     4921 gctgctggtg ccaaccctat tctccttctt cgccaaaagc actctgattg acaaatacga
     4981 tttatctaat ttacacgaaa ttgcttctgg tggcgctccc ctctctaagg aagtcgggga
     5041 agcggttgcc aagaggttcc atctgccagg tatcaggcaa ggatatgggc tcactgagac
     5101 tacatcagct attctgatta cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt
     5161 tgttccattt tttgaagcga aggttgtgga tctggatacc gggaaaacgc tgggcgttaa
     5221 tcaaagaggc gaactgtgtg tgagaggtcc tatgattatg tccggttatg taaacaatcc
     5281 ggaagcgacc aacgccttga ttgacaagga tggatggcta cattctggag acatagctta
     5341 ctgggacgaa gacgaacact tcttcatcgt tgaccgcctg aagtctctga ttaagtacaa
     5401 aggctatcag gtggctcccg ctgaattgga atccatcttg ctccaacacc ccaacatctt
     5461 cgacgcaggt gtcgcaggtc ttcccgacga tgacgccggt gaacttcccg ccgccgttgt
     5521 tgttttggag cacggaaaga cgatgacgga aaaagagatc gtggattacg tcgccagtca
     5581 agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt gtggacgaag taccgaaagg
     5641 tcttaccgga aaactcgacg caagaaaaat cagagagatc ctcataaagg ccaagaaggg
     5701 cggaaagatc gccgtgtaat tctagagtcg gggcggccgg ccgcttcgag cagacatgat
     5761 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     5821 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     5881 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     5941 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatccgt cgaccgatgc
     6001 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     6061 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     6121 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6181 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6241 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6301 ttccata
//
