LOCUS       sequence-399744-        6765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pGL3 luciferase reporter with JAM2 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 6765)
  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 6765)
  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-JAM2-SVA
FEATURES             Location/Qualifiers
     source          1..6765
                     /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        4293..4489
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4340..4475
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      4519..4528
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4525..6177
                     /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(4591..4609)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    6218..6339
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6229..6248)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6283..6302
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6587..6604)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6758..6765,1..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 ggtacccagc cttgtcaaca cctttatttc
     2521 agccttgtga tactttgggc agatacccag tcttcttgtt ggacttccgc tctacagaac
     2581 tatttagttg ttatgtaaca ctagaaaatg aataaactgg ccggatctgc tgcaaggttt
     2641 tctgatttaa atttctccta tactagtgca tcactagtat ttcagaagga gcatgaacaa
     2701 catgaaaaag gaagggggct ccctctccct ctccccctcc tcctccctct ccctccctct
     2761 ctgtctccgt ctccccacgg tctccctctg atgccgagcg gaagctggac tgtactgctg
     2821 ccatctcggc tcactgcaac ctccctgcct gattctcctg cctcagcctg ccgagcgcct
     2881 gggattgcag gcgcgcgccg ccacgcctga ctggttttcg tatttttttg gtggagacgg
     2941 ggtttcgcta tgttggccag gctggtctcc agctcctaac cgcgagtgat ctgccagcct
     3001 cggcctcccg aggtgccggg attgcagacg gagtctcgtt cactcagtgc tcagtgttgc
     3061 ccaggctgga gtgcagtggc gtgatctcgg ctcgctacaa cctccacctc ccagccgcct
     3121 gccttggcct cccaaagtgc cgagattgca gcctctgcct ggccgccacc ccgtctggga
     3181 agtgaggagc gtctctgcct ggccacccat cgtctgggat gtgaggagcc cctctgcctg
     3241 gctgcccagt ctgggaagtg aggagcgcct cttcccggcc gcccatcttc tgagatgtgg
     3301 gaagcgcctc tgccccgccg cccggtctgg gatgtgagga gcgcctctgc ccggccgcga
     3361 ccccgtctgg gaggtgagga gcgtctctgc ccggccaccc cgtctgagaa gtgaggagcc
     3421 cctccgcctg gcagccgccc catctggaaa gtgaggagcc cctccgcccg gcagccgccc
     3481 cgccccgtct gggaagtgag gagcatctcc gccctgccgc cgccccatcc cggagggagg
     3541 tgggggggcg cctctgcccg gccgccgccc cgtccgggag gtggggggcc cctctgcccg
     3601 gccgccctgt ctgggaagtg aggagcccct ctgccccgcg gccgccccgt ctgggaggtg
     3661 tacccaacag ctcattgaga acgggccatg atgacgatgg cggttttgtc gaatagaaaa
     3721 gggggaaatg tggggaaaag atagagaaat cagattgttg ctgtgtctgt gtagaaagta
     3781 gacataggag actccatttt gttctgtact aagaaaaatt cttctgcctt gggatgctgt
     3841 taatctataa cattaccccc aaccccgtgc tctctgaaac gtgtgctgtg tccactcagg
     3901 gttaaatgga ttaagggcgg tgcaagatgt gctttgttaa acagatgctt gaaggcagca
     3961 tgctccttaa gagtcatcac cactccctaa tctcaagtac ccagggacac aaacactgcg
     4021 gaaggccgca gggtcctctg cctaggaaaa ccaaagaccc ttgttcactt gtttatctgc
     4081 tgaccttccc tccactattg tcctatgacc ctgccaaatc cccctctgcg agaaacaccc
     4141 aagaatgatc aataaatact aaaaaaaaaa aaaagaaaga aaaaggaagg ggagctgttt
     4201 tagtcttgta acttggtaga ttatacacca ccaatgaaag agaagagatc gtgttcctcc
     4261 acaaagctag cccgggctcg agatctgcga tctgcatctc aattagtcag caaccatagt
     4321 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4381 ccatcgctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4441 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttggcat
     4501 tccggtactg ttggtaaagc caccatggaa gacgccaaaa acataaagaa aggcccggcg
     4561 ccattctatc cgctggaaga tggaaccgct ggagagcaac tgcataaggc tatgaagaga
     4621 tacgccctgg ttcctggaac aattgctttt acagatgcac atatcgaggt ggacatcact
     4681 tacgctgagt acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg atatgggctg
     4741 aatacaaatc acagaatcgt cgtatgcagt gaaaactctc ttcaattctt tatgccggtg
     4801 ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga acgacattta taatgaacgt
     4861 gaattgctca acagtatggg catttcgcag cctaccgtgg tgttcgtttc caaaaagggg
     4921 ttgcaaaaaa ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat tattatcatg
     4981 gattctaaaa cggattacca gggatttcag tcgatgtaca cgttcgtcac atctcatcta
     5041 cctcccggtt ttaatgaata cgattttgtg ccagagtcct tcgataggga caagacaatt
     5101 gcactgatca tgaactcctc tggatctact ggtctgccta aaggtgtcgc tctgcctcat
     5161 agaactgcct gcgtgagatt ctcgcatgcc agagatccta tttttggcaa tcaaatcatt
     5221 ccggatactg cgattttaag tgttgttcca ttccatcacg gttttggaat gtttactaca
     5281 ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt atagatttga agaagagctg
     5341 tttctgagga gccttcagga ttacaagatt caaagtgcgc tgctggtgcc aaccctattc
     5401 tccttcttcg ccaaaagcac tctgattgac aaatacgatt tatctaattt acacgaaatt
     5461 gcttctggtg gcgctcccct ctctaaggaa gtcggggaag cggttgccaa gaggttccat
     5521 ctgccaggta tcaggcaagg atatgggctc actgagacta catcagctat tctgattaca
     5581 cccgaggggg atgataaacc gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag
     5641 gttgtggatc tggataccgg gaaaacgctg ggcgttaatc aaagaggcga actgtgtgtg
     5701 agaggtccta tgattatgtc cggttatgta aacaatccgg aagcgaccaa cgccttgatt
     5761 gacaaggatg gatggctaca ttctggagac atagcttact gggacgaaga cgaacacttc
     5821 ttcatcgttg accgcctgaa gtctctgatt aagtacaaag gctatcaggt ggctcccgct
     5881 gaattggaat ccatcttgct ccaacacccc aacatcttcg acgcaggtgt cgcaggtctt
     5941 cccgacgatg acgccggtga acttcccgcc gccgttgttg ttttggagca cggaaagacg
     6001 atgacggaaa aagagatcgt ggattacgtc gccagtcaag taacaaccgc gaaaaagttg
     6061 cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca
     6121 agaaaaatca gagagatcct cataaaggcc aagaagggcg gaaagatcgc cgtgtaattc
     6181 tagagtcggg gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     6241 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     6301 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     6361 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     6421 aaatgtggta aaatcgataa ggatccgtcg accgatgccc ttgagagcct tcaacccagt
     6481 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
     6541 tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg ctcactgact
     6601 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6661 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6721 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccata
//
