LOCUS       sequence-12044-e        6154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for nuclear expression of small RNAs in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6154)
  AUTHORS   Good PD, Krikos AJ, Li SX, Bertrand E, Lee NS, Giver L, Ellington A,
            Zaia JA, Rossi JJ, Engelke DR
  TITLE     Expression of small, therapeutic RNAs in human cell nuclei.
  JOURNAL   Gene Ther. 1997 Jan . 4(1):45-54.
  PUBMED    9068795
REFERENCE   2  (bases 1 to 6154)
  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: "Gene Ther. 
            1997 Jan . 4(1):45-54."
COMMENT     Sequence Label: pAV U6+27
FEATURES             Location/Qualifiers
     source          1..6154
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     605..624
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1035..1365)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1049..1184
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1350..1370
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1398..1646
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1398..1418
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1569..1588
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    1793..1914
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   1963..2092
                     /label=AAV2 ITR
     rep_origin      complement(2223..2651)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2333..2354)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2545..2564
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2678..2782
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2783..2851,2852..3643)
                     /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     complement(3001..3020)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3814..4402
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4303..4322
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4845..4979
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5404..5424)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          5554..5619
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     primer_bind     complement(join(6151..6154,1..16))
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
       61 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
      121 gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct
      181 ggcgcgagcc cctgatgctc ttgtccagat catcctgatc gacaagaccg gcttccatcc
      241 gagtacgtgc tcgctcgatg cgatgttcgc ttggtggtcg aatgggcagg tagccggatc
      301 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
      361 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
      421 ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag
      481 ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
      541 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
      601 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
      661 tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag atcttgatcc
      721 cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc agggcttccc
      781 aaccttacca gagggcgccc cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc
      841 ccagtctagc tatcggcatg taagcccact gcaagctacc tgctttctct ttgcgcttgc
      901 gttttccctt gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg
      961 cggactggct ttctacgtgt tccgcttcct ttagcagccc ttgcgccctg agtgcttgcg
     1021 gcacggtgaa gctttttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg
     1081 aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca
     1141 tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc
     1201 gggactatgg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     1261 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct
     1321 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagccggat
     1381 ccaaggtcgg gcaggaagag ggcctatttc ccatgattcc ttcatatttg catatacgat
     1441 acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag atattagtac
     1501 aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt
     1561 ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat
     1621 atatcttgtg gaaaggacga aacaccgtgc tcgcttcggc agcacatata ctagtcgacg
     1681 ggtctagagc ggacttcggt ccgcttttta ctaggacctg caggcatgca agcttgacgt
     1741 cggttaccga tatccatatg gcggccgcat cgatctcgag ccgcggacta gtaacttgtt
     1801 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     1861 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     1921 cttacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg
     1981 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     2041 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agagagggag
     2101 tggccaaaga tctctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     2161 tgcgcagcct gaatggctaa tgggaaattg taaacgttaa tattttgtta atattttgtt
     2221 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2281 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2341 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2401 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2461 ccgtaaagca ctaaatcgga accctaaagg gatgccccga tttagagctt gacggggaaa
     2521 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     2581 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     2641 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2701 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2761 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2821 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2881 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2941 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3001 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3061 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3121 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3181 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3241 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3301 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3361 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3421 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3481 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3541 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3601 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3661 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3721 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3781 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3841 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3901 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3961 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4021 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4081 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4141 ttcgtgcaac acagccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4201 tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4261 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4321 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4381 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4441 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4501 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4561 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     4621 gccgattcat taatgcagag atctttggcc actccctctc tgcgcgctcg ctcgctcact
     4681 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     4741 gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg gaggggtgga
     4801 gtcgtgacgt gaattacgtc atagggttag ggaggtcctg gatcgatcca gacatgataa
     4861 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     4921 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     4981 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     5041 aaagcaagta aaacctctac aaatgtggta tggctgatta tgatctctag tcaaggcact
     5101 atacatcaaa tattccttat taaccccttt acaaattaaa aagctaaagg tacacaattt
     5161 ttgagcatag ttattaatag cagacactct atgcctgtgt ggagtaagaa aaaacagtat
     5221 gttatgatta taactgttat gcctacttat aaaggttaca gaatattttt ccataatttt
     5281 cttgtatagc agtgcagctt tttcctttgt ggtgtaaata gcaaagcaag caagagttct
     5341 attactaaac acagcatgac tcaaaaaact tagcaattct gaaggaaagt ccttggggtc
     5401 ttctaccttt ctcttctttt ttggaggagt agaatgttga gagtcagcag tagcctcatc
     5461 atcactagat ggcatttctt ctgagcaaaa caggttttcc tcattaaagg cattccacca
     5521 ctgctcccat tcatcagttc cataggttgg aatctaaaat acacaaacaa ttagaatcag
     5581 tagtttaaca cattatacac ttaaaaattt tatatttacc ttagagcttt aaatctctgt
     5641 aggtagtttg tccaattatg tcacaccaca gaagtaaggt tccttcacaa agatccggga
     5701 ccaaagcggc catcgtgcct ccccactcct gcagttcggg ggcatggatg cgcggatagc
     5761 cgctgctggt ttcctggatg ccgacggatt tgcactgccg gtagaactcc gcgaggtcgt
     5821 ccagcctcag gcagcagctg aaccaactcg cgaggggatc gagcccgggg tgggcgaaga
     5881 actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc
     5941 cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg
     6001 gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct cagaagaact cgtcaagaag
     6061 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg
     6121 gtcagcccat tcgccgccaa gctcttcagc aata
//
