LOCUS       40924_305        6741 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector ATG-less TV, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6741)
  AUTHORS   Konishi Y, Karnan S, Takahashi M, Ota A, Damdindorj L, Hosokawa Y, 
            Konishi H.
  TITLE     A system for the measurement of gene targeting efficiency in human 
            cell lines using an antibiotic resistance-GFP fusion gene
  JOURNAL   BioTechniques 53 (3), 141-152 (2012)
  PUBMED    22963476
REFERENCE   2  (bases 1 to 6741)
  AUTHORS   Konishi H.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-JAN-2012) Aichi Medical University School of Medicine,
            1-1 Yazako-Karimta, Nagakute, Aichi 480-1195, Japan
REFERENCE   3  (bases 1 to 6741)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6741)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "BioTechniques"; date: "2012"; volume: "53"; issue: "3"; pages: 
            "141-152"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (07-JAN-2012) Aichi Medical University School of Medicine, 1-1 
            Yazako-Karimta, Nagakute, Aichi 480-1195, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6741
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     CDS             180..896
                     /label=EGFP
                     /note="enhanced GFP"
     primer_bind     900..916
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    918..951
                     /label=loxP
                     /note="loxP"
     protein_bind    918..951
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     intron          989..1218
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     misc_feature    1259..1809
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             1823..2614
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     polyA_signal    complement(2639..2773)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(2870..2903)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2956..2972)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2975..3993
                     /note="partial SV40 largeT; an internal portion of SV40
                     largeT exon 2"
     repeat_region   4004..4144
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4219..4674
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4956..5060
                     /label=AmpR promoter
     CDS             5061..5918
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      6092..6680
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgag ctcgctagca cgcgtgcggc cgctctagat
      181 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
      241 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
      301 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
      361 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
      421 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
      481 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
      541 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
      601 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
      661 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
      721 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
      781 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
      841 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat
      901 ctagaactag tggatccata acttcgtata gcatacatta tacgaagtta tggaattaat
      961 tcgctgtctg cgagggccag ctgttggggt gagtactccc tctcaaaagc gggcatgact
     1021 tctgcgctaa gattgtcagt ttccaaaaac gaggaggatt tgatattcac ctggcccgcg
     1081 gtgatgcctt tgagggtggc cgcgtccatc tggtcagaaa agacaatctt tttgttgtca
     1141 agcttgaggt gtggcaggct tgagatctgg ccatacactt gagtgacaat gacatccact
     1201 ttgcctttct ctccacaggt gtccactccc aggtccaact gcaggtcgag cgtcgagaac
     1261 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     1321 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     1381 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     1441 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     1501 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     1561 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     1621 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     1681 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     1741 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     1801 cgatgataag cttgccacaa ccatggctga acaagatgga ttgcacgcag gttctccggc
     1861 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     1921 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     1981 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     2041 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     2101 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     2161 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     2221 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     2281 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     2341 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     2401 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     2461 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     2521 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     2581 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcggggga
     2641 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2701 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2761 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     2821 gtgggaggtt ttttctaggg ggccccggcg aggccgggat gagttgggaa taacttcgta
     2881 tagcatacat tatacgaagt tatccaaatt ttaaggtacc actgtgcata tgtatgaatt
     2941 cgatatcaag cttatcgata ccgtcgacct cgagcacaca ggcatagagt gtctgctatt
     3001 aataactatg ctcaaaaatt gtgtaccttt agctttttaa tttgtaaagg ggttaataag
     3061 gaatatttga tgtatagtgc cttgactaga gatccatttt ctgttattga ggaaagtttg
     3121 ccaggtgggt taaaggagca tgattttaat ccagaagaag cagaggaaac taaacaagtg
     3181 tcctggaagc ttgtaacaga gtatgcaatg gaaacaaaat gtgatgatgt gttgttattg
     3241 cttgggatgt acttggaatt tcagtacagt tttgaaatgt gtttaaaatg tattaaaaaa
     3301 gaacagccca gccactataa gtaccatgaa aagcattatg caaatgctgc tatatttgct
     3361 gacagcaaaa accaaaaaac catatgccaa caggctgttg atactgtttt agctaaaaag
     3421 cgggttgata gcctacaatt aactagagaa caaatgttaa caaacagatt taatgatctt
     3481 ttggatagga tggatataat gtttggttct acaggctctg ctgacataga agaatggatg
     3541 gctggagttg cttggctaca ctgtttgttg cccaaaatgg attcagtggt gtatgacttt
     3601 ttaaaatgca tggtgtacaa cattcctaaa aaaagatact ggctgtttaa aggaccaatt
     3661 gatagtggta aaactacatt agcagctgct ttgcttgaat tatgtggggg gaaagcttta
     3721 aatgttaatt tgcccttgga caggctgaac tttgagctag gagtagctat tgaccagttt
     3781 ttagtagttt ttgaggatgt aaagggcact ggaggggagt ccagagattt gccttcaggt
     3841 cagggaatta ataacctgga caatttaagg gattatttgg atggcagtgt taaggtaaac
     3901 ttagaaaaga aacacctaaa taaaagaact caaatatttc cccctggaat agtcaccatg
     3961 aatgagtaca gtgtgcctaa aacactgcag gcctcgaggc cgcaggaacc cctagtgatg
     4021 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     4081 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     4141 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4201 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4261 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4321 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4381 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     4441 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4501 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4561 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4621 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     4681 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     4741 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4801 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4861 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     4921 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4981 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5041 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     5101 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5161 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5221 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5281 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5341 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5401 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5461 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5521 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5581 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5641 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5701 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5761 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5821 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5881 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5941 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6001 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6061 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6121 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6181 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     6241 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6301 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6361 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6421 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6481 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6541 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6601 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6661 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6721 gctggccttt tgctcacatg t
//
