LOCUS       sequence-10124-e        5346 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 5346)
  AUTHORS   King GD, Muhammad AK, Xiong W, Kroeger KM, Puntel M, Larocque D, 
            Palmer D, Ng P, Lowenstein PR, Castro MG
  TITLE     High-capacity adenovirus vector-mediated anti-glioma gene therapy in
            the presence of systemic antiadenovirus immunity.
  JOURNAL   J Virol. 2008 May . 82(9):4680-4.
  PUBMED    18287240
REFERENCE   2  (bases 1 to 5346)
  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 Virol. 
            2008 May . 82(9):4680-4."
COMMENT     Sequence Label: pBS-MCS1M(1)Kana-mCMV-TK-pA
FEATURES             Location/Qualifiers
     source          1..5346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     65..84
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..320
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     312..329
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     313..329
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     339..358
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        339..357
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1248..1783
                     /label=mCMV promoter
                     /note="mouse cytomegalovirus (CMV) immediate early 
                     promoter"
     polyA_signal    1825..1959
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1862..1881)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1916..1935
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1979..1998)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1990..2805
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             1990..2805
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2063..2082)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     complement(2871..2891)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2871..2889)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2910..2926)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2910..2926)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2923..2945)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2934..2950
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2958..2964,2965..2982,2983..2988))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3003..3024
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3141..3158)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3312..3900)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3392..3411)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4071..4862,4863..4931))
                     /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     4694..4713
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4932..5036)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5062..5346,1..171))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5199..5220)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
       61 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
      121 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
      181 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
      241 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
      301 agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg
      361 aattgggtac cctagggcgg ccgcaagctt gaattcgagc tcgcccaact ccgcccgttt
      421 tatgactaga accaatagtt tttaatgcca aatgcactga aatcccctaa tttgcaaagc
      481 caaacgcccc ctatgtgagt aatacgggga ctttttaccc aatttcccaa gcggaaagcc
      541 ccctaataca ctcatatggc atatgaatca gcacggtcat gcactctaat ggcggcccat
      601 agggactttc cacatagggg gcgttcacca tttcccagca taggggtggt gactcaatgg
      661 cctttaccca agtacattgg gtcaatggga ggtaagccaa tgggtttttc ccattactgg
      721 caagcacact gagtcaaatg ggactttcca ctgggttttg cccaagtaca ttgggtcaat
      781 gggaggtgag ccaatgggaa aaacccattg ctgccaagta cactgactca atagggactt
      841 tccaatgggt ttttccattg ttggcaagca tataaggtca atgtgggtga gtcaataggg
      901 actttccatt gtattctgcc cagtacataa ggtcaatagg gggtgaatca acaggaaagt
      961 cccattggag ccaagtacac tgcgtcaata gggactttcc attgggtttt gcccagtaca
     1021 taaggtcaat aggggatgag tcaatgggaa aaacccattg gagccaagta cactgactca
     1081 atagggactt tccattgggt tttgcccagt acataaggtc aatagggggt gagtcaacag
     1141 gaaagtccca ttggagccaa gtacattgag tcaataggga ctttccaatg ggttttgccc
     1201 agtacataag gtcaatggga ggtaagccaa tgggtttttc ccattactgg cacgtatact
     1261 gagtcattag ggactttcca atgggttttg cccagtacat aaggtcaata ggggtgaatc
     1321 aacaggaaag tcccattgga gccaagtaca ctgagtcaat agggactttc cattgggttt
     1381 tgcccagtac aaaaggtcaa tagggggtga gtcaatgggt ttttcccatt attggcacgt
     1441 acataaggtc aataggggtg agtcattggg tttttccagc caatttaatt aaaacgccat
     1501 gtactttccc accattgacg tcaatgggct attgaaacta atgcaacgtg acctttaaac
     1561 ggtactttcc catagctgat taatgggaaa gtaccgttct cgagccaata cacgtcaatg
     1621 ggaagtgaaa gggcagccaa aacgtaacac cgccccggtt ttcccctgga aattccatat
     1681 tggcacgcat tctattggct gagctgcgtt ctacgtgggt ataagaggcg cgaccagcgt
     1741 cggtaccgtc gcagtcttcg gtctgaccac cgtagaacgc agagctcctc gctgcagccc
     1801 gggtctagag gatccgtcga cgttaacttg tttattgcag cttataatgg ttacaaataa
     1861 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     1921 ttgtccaaac tcatcaatgt atcttatcat gtctggatcc aagcttgcta gcgtaataca
     1981 aggggtgtta tgagccatat tcaacgggaa acgtcttgct ctaggccgcg attaaattcc
     2041 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     2101 gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     2161 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     2221 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     2281 accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc tgattcaggt
     2341 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     2401 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     2461 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     2521 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     2581 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     2641 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     2701 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     2761 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaagaatt aattcatgag
     2821 cggatacata tttgaatggc tagcgcggcc gcgagctcca gcttttgttc cctttagtga
     2881 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     2941 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3001 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3061 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3121 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3181 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3241 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3301 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3361 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3421 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3481 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3541 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3601 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3661 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3721 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     3781 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3841 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3901 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3961 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4021 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4081 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4141 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4201 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4261 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4321 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4381 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4441 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4501 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4561 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4621 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4681 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4741 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4801 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4861 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4921 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4981 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5041 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     5101 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     5161 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     5221 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     5281 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     5341 atcgga
//
