LOCUS       sequence-399743-        7116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pGL3 luciferase reporter with POFUT2 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 7116)
  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 7116)
  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-POFUT2-SVA
FEATURES             Location/Qualifiers
     source          1..7116
                     /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        4644..4840
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4691..4826
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      4870..4879
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4876..6528
                     /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(4942..4960)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    6569..6690
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6580..6599)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6634..6653
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6938..6955)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(7109..7116,1..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 ggtaccggat cctccagtaa ggacagatac
     2521 gtgactcttt agcagacaga catgcgtact tggcttctgg ttaattcgtt aggaactggc
     2581 tccaaaggtg cagacacaca actcagcttt accctcccag gaattacagt tgaagatcat
     2641 gcagatcttc atggatctgg gtagttccag gtctacccag aacagaacca gggatcaaga
     2701 agaaaggaaa gaaacgtgct ccctctccct ctccttctcc ctctccctct ccctctcccc
     2761 acggtctccc tctctttcca cggtctccct ctcatgcgga gccaaagctg gactgtgctg
     2821 ctgccatctc ggctcactgc aacctccctg cctggttctc ctgcctcagc ctgccgagtg
     2881 cctgcgattg caggcacgcg ccaccacgcc tgactggttt tggtggagac ggggtttcgc
     2941 tgtgttggcc gggccggtct ccagccccta accgcgagtg atccgccagc ctcggcctcc
     3001 cgaggtgctg ggattgcaga cggagtctcg ttcactcagt gctcgatggt gcccaggctg
     3061 gagtgcagtg gcgtgatctc ggctcgctac aacctacacc tcccagccgc ctgccttggc
     3121 ctcccaaagt gccaagattg cagcctctgc ccggccgcca ccccgtctgg gaagtgagga
     3181 gtgtctctgc ctggccgccc atcgtctggg atgtgaggag cccctctgcc tggctgccca
     3241 gtctggaaag tgaggagcat ctccgcccgg ccgccatccc atctaggaag tgaggagcac
     3301 ctctttccgg ccgccatcac atctaggaag tgaggagcgt ctctgcccag ccgcccatcg
     3361 tctgagatgt ggggagcgcc tctgccccgc cgccccatct gggatgtgag gagcgcctct
     3421 gcctggccac gaccccgtct gggaggtgag gagcgtctct gcccggccgc cccgtctgag
     3481 aagtgaggag cctctccgcc cggcagccac cccatctggg aagtgaggag cgtctccgcc
     3541 cggcagccac cccgtccggg agggaggtgg gggggtcagc cccccgcccg gccagccgcc
     3601 ccgtccggga gggaggtggg ggggtcagca ccccggccgg ccagccgccc cgtccgggag
     3661 ggaggtgggg gggtcagccc cccgcctggc cagccgcccc gtccgggagg tgaggggcgc
     3721 ctctgcccgg ccgcccctac tgggaagcaa agagcccctc tgcccggcca gccgccccgt
     3781 ctgggaggga ggtggggggg tcagcccccc gcctggccag ccgccccgtc cgggaggtga
     3841 ggggcgcctc tgcccggccg cccctactgg gaagtgaaga gcccctctgc ccggccagcc
     3901 gccccgtctg ggagggaggt gggggggtca gccccccgcc cggccagccg ccccgtccgg
     3961 gaggtgaggg gcgcctctgc ccggccgccc ctactgggaa gtgaagagcc cctctgcccg
     4021 gccaccaccc cgtctgggag gggtacccaa cagctcattg agaacaggcc aggatgacaa
     4081 tggcggcttt gtggaataga aaggcgggaa aggtggggaa aagattgaga aatcggatgg
     4141 ttgccgtgtc tgtgtagaaa gaagtagaca tgggagactt ttcattttgt tctgcactaa
     4201 gaaaaattct tctgccttgg gatcctgttg atctgtgacc ttacccccaa ccctgtgctc
     4261 tctgaaacat gtgctgtgtc cactcagggt taaatggatt aagggcggta caagatgtgc
     4321 tttgttaaac agacgcttga aggcagcatg ctcgttaaga gtcatcacca atccctaatc
     4381 tcaagtaatc agggacacaa acactgcgga aggccgcagg gtcctctgcc taggaaaacc
     4441 agagaccttt gttcacttgt ttatctgctg accttccctc cactattgtt ccatgaccct
     4501 gccaaaatcc ccgtctgtga gaaacaccca agaattatca ataaaaaaaa taaattaaaa
     4561 gaaaggaaag aaacgaaagc attgaagctc catacccttt ctgacggcat ctgtgggcta
     4621 gcccgggctc gagatctgcg atctgcatct caattagtca gcaaccatag tcccgcccct
     4681 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatcgctg
     4741 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     4801 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttggca ttccggtact
     4861 gttggtaaag ccaccatgga agacgccaaa aacataaaga aaggcccggc gccattctat
     4921 ccgctggaag atggaaccgc tggagagcaa ctgcataagg ctatgaagag atacgccctg
     4981 gttcctggaa caattgcttt tacagatgca catatcgagg tggacatcac ttacgctgag
     5041 tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac gatatgggct gaatacaaat
     5101 cacagaatcg tcgtatgcag tgaaaactct cttcaattct ttatgccggt gttgggcgcg
     5161 ttatttatcg gagttgcagt tgcgcccgcg aacgacattt ataatgaacg tgaattgctc
     5221 aacagtatgg gcatttcgca gcctaccgtg gtgttcgttt ccaaaaaggg gttgcaaaaa
     5281 attttgaacg tgcaaaaaaa gctcccaatc atccaaaaaa ttattatcat ggattctaaa
     5341 acggattacc agggatttca gtcgatgtac acgttcgtca catctcatct acctcccggt
     5401 tttaatgaat acgattttgt gccagagtcc ttcgataggg acaagacaat tgcactgatc
     5461 atgaactcct ctggatctac tggtctgcct aaaggtgtcg ctctgcctca tagaactgcc
     5521 tgcgtgagat tctcgcatgc cagagatcct atttttggca atcaaatcat tccggatact
     5581 gcgattttaa gtgttgttcc attccatcac ggttttggaa tgtttactac actcggatat
     5641 ttgatatgtg gatttcgagt cgtcttaatg tatagatttg aagaagagct gtttctgagg
     5701 agccttcagg attacaagat tcaaagtgcg ctgctggtgc caaccctatt ctccttcttc
     5761 gccaaaagca ctctgattga caaatacgat ttatctaatt tacacgaaat tgcttctggt
     5821 ggcgctcccc tctctaagga agtcggggaa gcggttgcca agaggttcca tctgccaggt
     5881 atcaggcaag gatatgggct cactgagact acatcagcta ttctgattac acccgagggg
     5941 gatgataaac cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat
     6001 ctggataccg ggaaaacgct gggcgttaat caaagaggcg aactgtgtgt gagaggtcct
     6061 atgattatgt ccggttatgt aaacaatccg gaagcgacca acgccttgat tgacaaggat
     6121 ggatggctac attctggaga catagcttac tgggacgaag acgaacactt cttcatcgtt
     6181 gaccgcctga agtctctgat taagtacaaa ggctatcagg tggctcccgc tgaattggaa
     6241 tccatcttgc tccaacaccc caacatcttc gacgcaggtg tcgcaggtct tcccgacgat
     6301 gacgccggtg aacttcccgc cgccgttgtt gttttggagc acggaaagac gatgacggaa
     6361 aaagagatcg tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga
     6421 gttgtgtttg tggacgaagt accgaaaggt cttaccggaa aactcgacgc aagaaaaatc
     6481 agagagatcc tcataaaggc caagaagggc ggaaagatcg ccgtgtaatt ctagagtcgg
     6541 ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     6601 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     6661 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     6721 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     6781 aaaatcgata aggatccgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt
     6841 ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca
     6901 actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6961 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7021 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7081 accgtaaaaa ggccgcgttg ctggcgtttt tccata
//
