LOCUS       sequence-263315-        6967 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 6967)
  TITLE     pShuttle vectors
REFERENCE   2  (bases 1 to 6967)
  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"
COMMENT     Sequence Label: U6 pShuttle
FEATURES             Location/Qualifiers
     source          1..6967
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..103
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     191..341
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
     promoter        390..638
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     390..410
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     561..580
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     repeat_region   3926..4028
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     primer_bind     complement(4117..4134)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4288..4876)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4368..4387)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             5708..6502
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5708..6502
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5762..5781)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6372..6391
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(6550..6930)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6612..6633)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6824..6843
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 catcatcaat aatatacctt attttggatt gaagccaata tgataatgag ggggtggagt
       61 ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg tagtagtgtg gcggaagtgt
      121 gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg tggcaaaagt gacgtttttg
      181 gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg gttttaggcg gatgttgtag
      241 taaatttggg cgtaaccgag taagatttgg ccattttcgc gggaaaactg aataagagga
      301 agtgaaatct gaataatttt gtgttactca tagcgcgtaa tactggtacc gcggccgcct
      361 cgacgcgtgg atccaaggtc gggcaggaag agggcctatt tcccatgatt ccttcatatt
      421 tgcatatacg atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa
      481 agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt
      541 taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt
      601 tcttggcttt atatatcttg tggaaaggac gaaacaccgt gctcgcttcg gcagcacata
      661 tactagtcga cagtactggt acctctagag cggacttcgg tccgcttttt actaggacct
      721 gcaggcatgc aagcttggga gatctgggcg tggttaaggg tgggaaagaa tatataaggt
      781 gggggtctta tgtagttttg tatctgtttt gcagcagccg ccgccgccat gagcaccaac
      841 tcgtttgatg gaagcattgt gagctcatat ttgacaacgc gcatgccccc atgggccggg
      901 gtgcgtcaga atgtgatggg ctccagcatt gatggtcgcc ccgtcctgcc cgcaaactct
      961 actaccttga cctacgagac cgtgtctgga acgccgttgg agactgcagc ctccgccgcc
     1021 gcttcagccg ctgcagccac cgcccgcggg attgtgactg actttgcttt cctgagcccg
     1081 cttgcaagca gtgcagcttc ccgttcatcc gcccgcgatg acaagttgac ggctcttttg
     1141 gcacaattgg attctttgac ccgggaactt aatgtcgttt ctcagcagct gttggatctg
     1201 cgccagcagg tttctgccct gaaggcttcc tcccctccca atgcggttta aaacataaat
     1261 aaaaaaccag actctgtttg gatttggatc aagcaagtgt cttgctgtct ttatttaggg
     1321 gttttgcgcg cgcggtaggc ccgggaccag cggtctcggt cgttgagggt cctgtgtatt
     1381 ttttccagga cgtggtaaag gtgactctgg atgttcagat acatgggcat aagcccgtct
     1441 ctggggtgga ggtagcacca ctgcagagct tcatgctgcg gggtggtgtt gtagatgatc
     1501 cagtcgtagc aggagcgctg ggcgtggtgc ctaaaaatgt ctttcagtag caagctgatt
     1561 gccaggggca ggcccttggt gtaagtgttt acaaagcggt taagctggga tgggtgcata
     1621 cgtggggata tgagatgcat cttggactgt atttttaggt tggctatgtt cccagccata
     1681 tccctccggg gattcatgtt gtgcagaacc accagcacag tgtatccggt gcacttggga
     1741 aatttgtcat gtagcttaga aggaaatgcg tggaagaact tggagacgcc cttgtgacct
     1801 ccaagatttt ccatgcattc gtccataatg atggcaatgg gcccacgggc ggcggcctgg
     1861 gcgaagatat ttctgggatc actaacgtca tagttgtgtt ccaggatgag atcgtcatag
     1921 gccattttta caaagcgcgg gcggagggtg ccagactgcg gtataatggt tccatccggc
     1981 ccaggggcgt agttaccctc acagatttgc atttcccacg ctttgagttc agatgggggg
     2041 atcatgtcta cctgcggggc gatgaagaaa acggtttccg gggtagggga gatcagctgg
     2101 gaagaaagca ggttcctgag cagctgcgac ttaccgcagc cggtgggccc gtaaatcaca
     2161 cctattaccg gctgcaactg gtagttaaga gagctgcagc tgccgtcatc cctgagcagg
     2221 ggggccactt cgttaagcat gtccctgact cgcatgtttt ccctgaccaa atccgccaga
     2281 aggcgctcgc cgcccagcga tagcagttct tgcaaggaag caaagttttt caacggtttg
     2341 agaccgtccg ccgtaggcat gcttttgagc gtttgaccaa gcagttccag gcggtcccac
     2401 agctcggtca cctgctctac ggcatctcga tccagcatat ctcctcgttt cgcgggttgg
     2461 ggcggctttc gctgtacggc agtagtcggt gctcgtccag acgggccagg gtcatgtctt
     2521 tccacgggcg cagggtcctc gtcagcgtag tctgggtcac ggtgaagggg tgcgctccgg
     2581 gctgcgcgct ggccagggtg cgcttgaggc tggtcctgct ggtgctgaag cgctgccggt
     2641 cttcgccctg cgcgtcggcc aggtagcatt tgaccatggt gtcatagtcc agcccctccg
     2701 cggcgtggcc cttggcgcgc agcttgccct tggaggaggc gccgcacgag gggcagtgca
     2761 gacttttgag ggcgtagagc ttgggcgcga gaaataccga ttccggggag taggcatccg
     2821 cgccgcaggc cccgcagacg gtctcgcatt ccacgagcca ggtgagctct ggccgttcgg
     2881 ggtcaaaaac caggtttccc ccatgctttt tgatgcgttt cttacctctg gtttccatga
     2941 gccggtgtcc acgctcggtg acgaaaaggc tgtccgtgtc cccgtataca gacttgagag
     3001 ggagtttaaa cgaattcaat agcttgttgc atgggcggcg atataaaatg caaggtgctg
     3061 ctcaaaaaat caggcaaagc ctcgcgcaaa aaagaaagca catcgtagtc atgctcatgc
     3121 agataaaggc aggtaagctc cggaaccacc acagaaaaag acaccatttt tctctcaaac
     3181 atgtctgcgg gtttctgcat aaacacaaaa taaaataaca aaaaaacatt taaacattag
     3241 aagcctgtct tacaacagga aaaacaaccc ttataagcat aagacggact acggccatgc
     3301 cggcgtgacc gtaaaaaaac tggtcaccgt gattaaaaag caccaccgac agctcctcgg
     3361 tcatgtccgg agtcataatg taagactcgg taaacacatc aggttgattc acatcggtca
     3421 gtgctaaaaa gcgaccgaaa tagcccgggg gaatacatac ccgcaggcgt agagacaaca
     3481 ttacagcccc cataggaggt ataacaaaat taataggaga gaaaaacaca taaacacctg
     3541 aaaaaccctc ctgcctaggc aaaatagcac cctcccgctc cagaacaaca tacagcgctt
     3601 ccacagcggc agccataaca gtcagcctta ccagtaaaaa agaaaaccta ttaaaaaaac
     3661 accactcgac acggcaccag ctcaatcagt cacagtgtaa aaaagggcca agtgcagagc
     3721 gagtatatat aggactaaaa aatgacgtaa cggttaaagt ccacaaaaaa cacccagaaa
     3781 accgcacgcg aacctacgcc cagaaacgaa agccaaaaaa cccacaactt cctcaaatcg
     3841 tcacttccgt tttcccacgt tacgtcactt cccattttaa gaaaactaca attcccaaca
     3901 catacaagtt actccgccct aaaacctacg tcacccgccc cgttcccacg ccccgcgcca
     3961 cgtcacaaac tccaccccct cattatcata ttggcttcaa tccaaaataa ggtatattat
     4021 tgatgatgtt aattaacatg catggatcca tatgcggtgt gaaataccgc acagatgcgt
     4081 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4141 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4201 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4261 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4321 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4381 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4441 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4501 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4561 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4621 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4681 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     4741 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4801 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4861 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4921 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4981 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5041 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5101 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5161 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5221 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5281 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5341 gcgcaacgtt gttgccattg ctgcagccat gagattatca aaaaggatct tcacctagat
     5401 ccttttcacg tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta
     5461 ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg
     5521 gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc
     5581 agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt
     5641 gccgccaagg atctgatggc gcaggggatc aagctctgat caagagacag gatgaggatc
     5701 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     5761 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     5821 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     5881 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     5941 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     6001 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     6061 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     6121 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     6181 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     6241 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     6301 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     6361 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     6421 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     6481 ccttcttgac gagttcttct gaattttgtt aaaatttttg ttaaatcagc tcatttttta
     6541 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     6601 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     6661 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     6721 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     6781 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     6841 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     6901 ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca ttcaggatcg aattaattct
     6961 taattaa
//
