LOCUS       sequence-10404-e        5763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5763)
  AUTHORS   Lau P, Verrier JD, Nielsen JA, Johnson KR, Notterpek L, Hudson LD
  TITLE     Identification of dynamically regulated microRNA and mRNA networks 
            in developing oligodendrocytes.
  JOURNAL   J Neurosci. 2008 Nov 5. 28(45):11720-30.
  PUBMED    18987208
REFERENCE   2  (bases 1 to 5763)
  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: "J Neurosci.
            2008 Nov 5. 28(45):11720-30."
COMMENT     Sequence Label: pcDNA6.2-EmGFP-mir9
FEATURES             Location/Qualifiers
     source          1..5763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        384..587
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     538..558
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        632..650
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    680..704
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(713..715,716..718,719..1432)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(758..779)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1019..1038)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1366..1387
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     ncRNA           1492..1518
                     /label=5' miR-155
                     /note="sequence upstream of the precursor of mouse miR-155 
                     microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     ncRNA           1586..1627
                     /label=3' miR-155
                     /note="sequence downstream of the precursor of mouse 
                     miR-155 microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     protein_bind    complement(1656..1680)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1722..1741)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1766..1814
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        complement(1985..2003)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2008..2024)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2008..2024)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      2092..2520
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2179..2198)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2389..2410
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2529..2549)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2534..2863
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2714..2849
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2776..2795
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2911..2958
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2977..3375
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3533..3654
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3570..3589)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3624..3643
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3689..3706)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3860..4448)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3940..3959)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4544..5335)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      421 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      481 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      541 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
      601 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagtcccaag
      661 ctggctagtt aagctatcaa caagtttgta caaaaaagca ggctttaaaa ccatggtgag
      721 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
      781 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
      841 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
      901 caccttcacc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
      961 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1021 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1081 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1141 gtacaactac aacagccaca aggtctatat caccgccgac aagcagaaga acggcatcaa
     1201 ggtgaacttc aagacccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     1261 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1321 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1381 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagctaagca
     1441 cttcgtggcc gtcgatcgtt taaagggagg tagtgagtcg accagtggat cctggaggct
     1501 tgctgaaggc tgtatgctgt ctttggttat ctagctgtat gagttttggc cactgactga
     1561 ctcatacaga gataaccaaa gacaggacac aaggcctgtt actagcactc acatggaaca
     1621 aatggcccag atctggccgc actcgagata tctagaccca gctttcttgt acaaagtggt
     1681 tgatctagag ggcccgcggt tcgctgatgg gggaggctaa ctgaaacacg gaaggagaca
     1741 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
     1801 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     1861 accgtgaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     1921 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     1981 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
     2041 tctcaaaatc tctgatggat ctgcgcagct ggggctctag ggggtatccc cacgcgccct
     2101 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2161 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2221 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2281 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2341 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2401 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2461 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2521 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2581 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2641 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2701 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2761 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     2821 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     2881 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc
     2941 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagccttt gtctcaagaa
     3001 gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac
     3061 tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta
     3121 tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg
     3181 gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc
     3241 ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg
     3301 aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat
     3361 gtgtgggagg gctaagcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt
     3421 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     3481 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     3541 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3601 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     3661 ctgtataccg tcgctcttcc gctgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3721 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3781 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3841 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3901 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3961 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4021 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4081 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4141 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4201 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4261 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4321 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4381 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4441 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgacgcgt
     4501 aactcacgtt aagggatttt ggtcatgggt ggctcgacga gggttatttg ccgactacct
     4561 tggtgatctc gcctttcacg tagtggacaa attcttccaa ctgatctgcg cgcgaggcca
     4621 agcgatcttc ttcttgtcca agataagcct gtctagcttc aagtatgacg ggctgatact
     4681 gggccggcag gcgctccatt gcccagtcgg cagcgacatc cttcggcgcg attttgccgg
     4741 ttactgcgct gtaccaaatg cgggacaacg taagcactac atttcgctca tcgccagccc
     4801 agtcgggcgg cgagttccat agcgttaagg tttcatttag cgcctcaaat agatcctgtt
     4861 caggaaccgg atcaaagagt tcctccgccg ctggacctac caaggcaacg ctatgttctc
     4921 ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt ggctggctcg aagatacctg
     4981 caagaatgtc attgcgctgc cattctccaa attgcagttc gcgcttagct ggataacgcc
     5041 acggaatgat gtcgtcgtgc acaacaatgg tgacttctac agcgcggaga atctcgctct
     5101 ctccagggga agccgaagtt tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat
     5161 caagccttac ggtcaccgta accagcaaat caatatcact gtgtggcttc aggccgccat
     5221 ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga
     5281 cgccaactac ctctgatagt tgagtcgata cttcggcgat caccgcttcc ctcataatgt
     5341 ttaactttgt tttagggcga ctgccctgct gcgtaacatc gttgctgctc cataacatca
     5401 aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata gactgtaccc
     5461 caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt
     5521 cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta cttgcattac agcttacgaa
     5581 ccgaacaggc ttatgtccac tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc
     5641 cggcaacctt gggtagcagc gaagtcgagg catttctgtc ctggctggtc tagaattgca
     5701 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
     5761 cgc
//
