LOCUS       Exported                4287 bp ds-DNA     circular SYN 19-JUL-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4287)
  AUTHORS   Jiten Doshi
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 4, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4287
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     226..245
                     /label=PR-JD37
                     /note="pJD02/03 sequencing primer"
     misc_feature    233..304
                     /label=upstream MCS
                     /note="upstream multiple cloning site"
     primer_bind     234..260
                     /label=Forward (CW) Analysis Primer
     promoter        254..272
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     254..271
                     /label=SP6
                     /note="Genewiz primer collection"
     misc_feature    326..336
                     /label=BsmBI insert
                     /note="fragment released by digestion with BsmBI"
     primer_bind     337..359
                     /label=JD105
                     /note="CMV-e1a(v1/v2)-BgHpA-cloning; 3 fragments; BsaI; CMV
                     promoter from pJD34, e1A-v1 gene from JD1-33-01 gene 
                     fragment or  e1A-v2 gene from JD1-33-02 gene fragment,, 
                     BgHpA from JD1-33-9 gene fragment"
     primer_bind     337..359
                     /label=PR-JD61
                     /note="CMV promoter amplify fp; template pJD34; for golden 
                     gate assembly of CMV-Cap-AAV9-gapdh-pA"
     primer_bind     337..359
                     /label=JD99
                     /note="CMV-e4orf6-BgHpA-cloning; 3 fragments; BbsI; CMV 
                     promoter from pJD34, e4orf6 gene from JD1-33-05 gene 
                     fragment, BgHpA from JD1-33-9 gene fragment"
     enhancer        341..644
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(416..441)
                     /label=JD149
                     /note="CMV rp for qPCR"
     promoter        645..848
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     663..687
                     /label=HL1169
                     /note="CMV seq"
     primer_bind     704..728
                     /label=HL1170
                     /note="CMV seq"
     primer_bind     799..819
                     /label=CMV-Forward
                     /note="Genewiz primer collection"
     primer_bind     complement(829..850)
                     /label=JD146
                     /note="rp; template pJD34; for part1 of CMV-E2A (mod5utr) 
                     cloning; part3 similar to pJD78; use JD105 as fp"
     primer_bind     complement(829..849)
                     /label=JD111
                     /note="CMV-e1b-BgHpA-cloning; 3 fragments; BsaI; CMV 
                     promoter amplification for golden gate assembly of E1B 
                     gene; reverse primer; use JD81 as forward primer; use 
                     JD85/86 for BghpA JD1-33-09 fragment"
     primer_bind     complement(829..848)
                     /label=JD106
     primer_bind     complement(829..848)
                     /label=JD114
                     /note="CMV-E2A BgHpA plasmid; BsaI golden gate; use JD85/86
                     for BghpA JD1-33-09 fragment; use JD81 as fp and JD90 as rp
                     for CMV promoter part; JD91/92 for E2A gene from 
                     pJDxx-E2A-Gapdh(pA)"
     primer_bind     complement(830..848)
                     /label=JD154
                     /note="pJD34 amp; CMV amp; part1 of GG of pJD97 
                     (cmv-e4orf6(mod5utr)-pA)"
     primer_bind     complement(831..852)
                     /label=JD100
     primer_bind     complement(831..848)
                     /label=PR-JD62
                     /note="CMV promoter amplify rp; template pJD34; for golden 
                     gate assembly of CMV-Cap-AAV9-gapdh-pA"
     misc_feature    849..914
                     /label=5UTR
     prim_transcript 849..909
                     /gene="E4"
                     /locus_tag="HAdVC_gp16"
                     /old_locus_tag="HAdVCgp31"
                     /experiment="transcript mapping"
     primer_bind     849..868
                     /label=JD136
                     /note="fp; template pE4orf6; e4orf6 gene in pJD07; dig pcr 
                     amp product with PacI, NotI; backbone pJD07 dig with PacI, 
                     NotI"
     primer_bind     849..868
                     /label=JD101
     prim_transcript 910..914
                     /gene="E4"
                     /locus_tag="HAdVC_gp16"
                     /old_locus_tag="HAdVCgp31"
                     /experiment="transcript mapping"
     CDS             join(915..1088,1800..2078)
                     /codon_start=1
                     /gene="E4"
                     /locus_tag="HAdVC_gp16"
                     /old_locus_tag="HAdVCgp31"
                     /product="control protein E4orf6/7"
                     /function="gene regulation;cell cycle regulation"
                     /label=E4orf6/7
                     /note="genus-specific"
                     /db_xref="GI:160409927"
                     /db_xref="GeneID:2652989"
                     /protein_id="YP_001551772.1"
                     /translation="MTTSGVPFGMTLRPTRSRLSRRTPYSRDRLPPFETETRATILEDH
                     PLLPECNTLTMHNAWTSPSPPVKQPQVGQQPVAQQLDSDMNLSELPGEFINITDERLAR
                     QETVWNITPKNMSVTHDMMLFKASRGERTVYSVCWEGGGRLNTRVL"
     CDS             915..1799
                     /codon_start=1
                     /product="E4 34K"
                     /label=E4orf6
                     /protein_id="AP_000228.1"
                     /translation="MTTSGVPFGMTLRPTRSRLSRRTPYSRDRLPPFETETRATILEDH
                     PLLPECNTLTMHNVSYVRGLPCSVGFTLIQEWVVPWDMVLTREELVILRKCMHVCLCCA
                     NIDIMTSMMIHGYESWALHCHCSSPGSLQCIAGGQVLASWFRMVVDGAMFNQRFIWYRE
                     VVNYNMPKEVMFMSSVFMRGRHLIYLRLWYDGHVGSVVPAMSFGYSALHCGILNNIVVL
                     CCSYCADLSEIRVRCCARRTRRLMLRAVRIIAEETTAMLYSCRTERRRQQFIRALLQHH
                     RPILMHDYDSTPM"
     gene            915..1675
                     /gene="E4"
                     /locus_tag="HAdVC_gp16"
                     /old_locus_tag="HAdVCgp31"
                     /label=E4
                     /db_xref="GeneID:2652989"
     prim_transcript 915..1675
                     /gene="E4"
                     /locus_tag="HAdVC_gp16"
                     /old_locus_tag="HAdVCgp31"
                     /experiment="transcript mapping"
     CDS             915..1675
                     /codon_start=1
                     /gene="E4"
                     /locus_tag="HAdVC_gp16"
                     /old_locus_tag="HAdVCgp31"
                     /product="control protein E4 34K"
                     /function="apoptosis;DNA damage response;RNA metabolism and
                     transport;transformation"
                     /label=E4
                     /note="E4orf6; genus-specific; 34K family"
                     /db_xref="GI:160409928"
                     /db_xref="GeneID:2652989"
                     /protein_id="NP_597755.2"
                     /translation="MTTSGVPFGMTLRPTRSRLSRRTPYSRDRLPPFETETRATILEDH
                     PLLPECNTLTMHNVSYVRGLPCSVGFTLIQEWVVPWDMVLTREELVILRKCMHVCLCCA
                     NIDIMTSMMIHGYESWALHCHCSSPGSLQCIAGGQVLASWFRMVVDGAMFNQRFIWYRE
                     VVNYNMPKEVMFMSSVFMRGRHLIYLRLWYDGHVGSVVPAMSFGYSALHCGILNNIVVL
                     CCSYCADLSEIRVRCCARRTRRLMLRAVRII"
     CDS             915..994
                     /codon_start=3
                     /gene="E4"
                     /locus_tag="HAdVC_gp16"
                     /old_locus_tag="HAdVCgp31"
                     /product="control protein E4orf4"
                     /function="RNA splicing"
                     /label=E4orf4
                     /note="genus-specific"
                     /db_xref="GI:160409929"
                     /db_xref="GeneID:2652989"
                     /protein_id="YP_001551773.1"
                     /translation="DYVRRSIWHDTTTNTISVVSAHSVQ"
     misc_feature    1109
                     /label=remove BbsI site
                     /note="/T to C silent base substitution"
     primer_bind     complement(1181..1197)
                     /label=JD144
                     /note="rp for cloning JD1-91-4 gene fragment cotaining 
                     E4orf6 partial gene sequence with 5utr, kozak and res. 
                     sites; fp JD72; "
     misc_feature    1646
                     /label=remove BsmBi site
                     /note="/T to G base substitution; silent"
     primer_bind     1674..1693
                     /label=JD255
     misc_feature    1676..2078
                     /label=E4-Region
                     /note="/vntifkey=21"
     CDS             1676..1799
                     /codon_start=1
                     /label=Orf-6
                     /note="/vntifkey=4"
                     /translation="EETTAMLYSCRTERRRQQFIRALLQHHRPILMHDYDSTPM"
     primer_bind     complement(1780..1799)
                     /label=JD102
     primer_bind     1794..1812
                     /label=JD185
                     /note="sequencing primer E4 region DeltaF6/pJD106,108 
                     colonyPCR"
     CDS             1800..2078
                     /codon_start=1
                     /product="E4 ORF6/7"
                     /label=E4 ORF6/7
                     /protein_id="AP_000227.1"
                     /translation="AWTSPSPPVKQPQVGQQPVAQQLDSDMNLSELPGEFINITDERLA
                     RQETVWNITPKNMSVTHDMMLFKASRGERTVYSVCWEGGGRLNTRVL"
     misc_feature    2079..2175
                     /label=3utr e4 gene
     primer_bind     complement(2116..2135)
                     /label=JD184
                     /note="sequencing primer E4 region DeltaF6"
     primer_bind     complement(2176..2213)
                     /label=JD254
     primer_bind     complement(2193..2212)
                     /label=JD178
                     /note="amplification of JD1-131-1 fragment for pJD106, 
                     pJD108 cloning"
     CDS             complement(2205..2207)
                     /codon_start=1
                     /product="fiber"
                     /label=fiber
                     /protein_id="AP_000226.1"
                     /translation=""
     misc_feature    2210..2238
                     /label=downstream MCS
                     /note="downstream multiple cloning site"
     primer_bind     complement(2215..2233)
                     /label=JD147
                     /note="colonyPCr/sequencing primer pJD02/03"
     primer_bind     complement(2243..2268)
                     /label=Reverse (CCW) Analysis Primer
     primer_bind     complement(2249..2268)
                     /label=T7
                     /note="Genewiz primer collection"
     promoter        complement(2250..2268)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2325..2335)
                     /label=PR-JD31
                     /note="pJD11 Ef1a-rtTA sequencing primer"
     primer_bind     2336..2355
                     /label=JD217
                     /note="fp2; frag2; gibson assembly; pJD83 template; pJD108 
                     backbone"
     promoter        2360..2464
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2408..2427)
                     /label=PR-JD31
                     /note="pJD11 Ef1a-rtTA sequencing primer"
     CDS             2465..3325
                     /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(2683..2702)
                     /label=Amp-R
                     /note="Quinatarabio5' end of ampiciltrn resistance gene, 
                     reverse primer"
     misc_feature    3326..3409
                     /label=terminator sequence?
     primer_bind     complement(3397..3415)
                     /label=PR-JD02
                     /note="RP to amplify gene fragment JD1-01 for AmpR gene 
                     cloning in golden gate backbone; dig by BanI, BsaAI"
     promoter        complement(3410..3422)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3487..3505)
                     /label=PR-JD32
                     /note="pJD02/03 sequencing primer"
     rep_origin      3523..4111
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3688..3711)
                     /label=HL1060
                     /note="pHL011 seq"
     primer_bind     3796..3815
                     /label=HL1044
                     /note="ori"
     primer_bind     4047..4066
                     /label=PR-JD29
                     /note="pJD11 rtTA CDS, pA sequencing primer"
     primer_bind     4221..4239
                     /label=PR-JD28
                     /note="pJD10 tTA CDS sequencing primer"
ORIGIN
        1 cgaaaaatca ataatcagac aacaagatgt gcgaactcga tattttacac gactctcttt
       61 accaattctg ccccgaatta cacttaaaac gactcaacag cttaacgttg gcttgccacg
      121 cattacttga ctgtaaaact ctcactctta ccgaacttgg ccgtaacctg ccaaccaaag
      181 cgagaacaaa acataacatc aaacgaatcg accgattgtt aggtaatcgt cacctgcagg
      241 aaggtttaaa cgcatttagg tgacactata gaagtgtgta tcgctcgagg gatccgaatt
      301 cgaagtcttg gtacggagcg agaccggagc gagacgggag cgttacataa cttacggtaa
      361 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      421 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      481 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      541 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      601 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      661 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      721 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      781 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      841 gcagagctca ctgtgactga ttgagctggt gccgtgtcga gtggtgtttt ttaataggtt
      901 tttttactgg aaatatgact acgtccggcg ttccatttgg catgacacta cgaccaacac
      961 gatctcggtt gtctcggcgc actccgtaca gtagggatcg cctacctcct tttgagacag
     1021 agacccgcgc taccatactg gaggatcatc cgctgctgcc cgaatgtaac actttgacaa
     1081 tgcacaacgt gagttacgtg cgaggtctcc cctgcagtgt gggatttacg ctgattcagg
     1141 aatgggttgt tccctgggat atggttctga cgcgggagga gcttgtaatc ctgaggaagt
     1201 gtatgcacgt gtgcctgtgt tgtgccaaca ttgatatcat gacgagcatg atgatccatg
     1261 gttacgagtc ctgggctctc cactgtcatt gttccagtcc cggttccctg cagtgcatag
     1321 ccggcgggca ggttttggcc agctggttta ggatggtggt ggatggcgcc atgtttaatc
     1381 agaggtttat atggtaccgg gaggtggtga attacaacat gccaaaagag gtaatgttta
     1441 tgtccagcgt gtttatgagg ggtcgccact taatctacct gcgcttgtgg tatgatggcc
     1501 acgtgggttc tgtggtcccc gccatgagct ttggatacag cgccttgcac tgtgggattt
     1561 tgaacaatat tgtggtgctg tgctgcagtt actgtgctga tttaagtgag atcagggtgc
     1621 gctgctgtgc ccggaggaca aggcggctca tgctgcgggc ggtgcgaatc atcgctgagg
     1681 agaccactgc catgttgtat tcctgcagga cggagcggcg gcggcagcag tttattcgcg
     1741 cgctgctgca gcaccaccgc cctatcctga tgcacgatta tgactctacc cccatgtagg
     1801 cgtggacttc tccttcgccg cccgttaagc aaccgcaagt tggacagcag cctgtggctc
     1861 agcagctgga cagcgacatg aacttaagtg agctgcccgg ggagtttatt aatatcactg
     1921 atgagcgttt ggctcgacag gaaaccgtgt ggaatataac acctaagaat atgtctgtta
     1981 cccatgatat gatgcttttt aaggccagcc ggggagaaag gactgtgtac tctgtgtgtt
     2041 gggagggagg tggcaggttg aatactaggg ttctgtgagt ttgattaagg tacggtgatc
     2101 tgtataagct atgtggtggt ggggctatac tactgaatga aaaatgactt gaaattttct
     2161 gcaattgaaa aataaacacg ttgaaacata acacaaacga ttctttaatt aagacgtcag
     2221 aattctcgag gcggccgcat gtgagtctcc ctatagtgag tcgtattaat ttcgcgggcg
     2281 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagtagca
     2341 ccaggcgttt aagggcaccc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2401 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2461 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2521 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2581 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2641 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     2701 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2761 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     2821 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     2881 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     2941 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3001 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3061 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3121 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3181 atctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3241 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3301 tgcgtcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3361 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt acgtgccgat
     3421 caaagtctca ttttcgccaa aagttgtcat gaccaaaatc ccttaacgtg agttttcgtt
     3481 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3541 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3601 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3661 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3721 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3781 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3841 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3901 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3961 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4021 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4081 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aatgcggcct ttttacggtt
     4141 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     4201 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     4261 gcgcagcgag tcagtgagcg aggaagc
//
