LOCUS Exported 5660 bp ds-DNA circular SYN 10-JAN-2022 DEFINITION Block 1 with deletion in E1 and insertion of GFP expression cassette. ACCESSION . VERSION . KEYWORDS pAd5-B1-deltaE1-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5660) TITLE AdenoBuilder JOURNAL Unpublished REFERENCE 2 (bases 1 to 5660) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 10, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5660 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(193..212) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 540..561 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(693..712) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 758..813 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(812..831) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(1174..1190) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1174..1190) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" protein_bind 1198..1214 /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(1914..1944) /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" protein_bind 1959..1980 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2100..2117) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2271..2859) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2351..2370) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3030..3890) /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 3653..3672 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3891..3995) /gene="bla" /label=AmpR promoter primer_bind 4063..4081 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind 4387..4406 /label=T7 /note="T7 promoter, forward primer" promoter 4387..4405 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" repeat_region 4461..4563 /label=ITR /note="inverted terminal repeat of human adenovirus serotype 5" misc_signal 4651..4801 /label=Ad5 Psi /note="packaging signal for adenovirus serotype 5" enhancer 5010..5313 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 5314..5517 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5468..5488 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 5541..5550 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 5547..606 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 5547..603 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(5592..5613) /label=EGFP-N /note="EGFP, reverse primer" ORIGIN 1 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg 61 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac 121 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc 181 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac 241 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg 301 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag 361 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag 421 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac 481 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac 541 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac 601 aagtaaagcg gccgcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg 661 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg 721 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt 781 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat 841 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg 901 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct 961 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt 1021 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga 1081 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc 1141 ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg 1201 ttatccgctc acaattccac acaacatacg agccggacta gttggcttaa gggtgggaaa 1261 gaatatataa ggtgggggtc ttatgtagtt ttgtatctgt tttgcagcag ccgccgccgc 1321 catgagcacc aactcgtttg atggaagcat tgtgagctca tatttgacaa cgcgcatgcc 1381 cccatgggcc ggggtgcgtc agaatgtgat gggctccagc attgatggtc gccccgtcct 1441 gcccgcaaac tctactacct tgacctacga gattcgaaca tctttctaga agatctccta 1501 caatattctc agctgccatg gaaaatcgat gttcttcttt tattctctca agattttcag 1561 gctgtatatt aaaacttata ttaagaacta tgctaaccac ctcatcagga accgttgtag 1621 gtggcgtggg ttttcttggc aatcgactct catgaaaact acgagctaaa tattcaatat 1681 gttcctcttg accaacttta ttctgcattt tttttgaacg aggtttagag caagcttcag 1741 gaaactgaga caggaatttt attaaaaatt taaattttga agaaagttca gggttaatag 1801 catccatttt ttgctttgca agttcctcag cattcttaac aaaagacgtc tcttttgaca 1861 tgtttaaagt ttaaacctcc tgtgtgaaat tattatccgc tcataattcc acacattata 1921 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 1981 attgcgttgc gctcactgcc aattgctttc cagtcgggaa acctgtcgtg ccagctgcat 2041 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 2101 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 2161 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 2221 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 2281 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 2341 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 2401 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 2461 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 2521 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 2581 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 2641 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 2701 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 2761 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 2821 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 2881 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 2941 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 3001 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 3061 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 3121 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 3181 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 3241 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 3301 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 3361 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 3421 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 3481 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 3541 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 3601 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 3661 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 3721 ctctcaagga tcttaccgct gttgagatcc agttcaatgt aacccactcg tgcacccaac 3781 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 3841 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 3901 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 3961 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 4021 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg 4081 ccgcccctgc agccgaatta tattattttt gccaaataat ttttaacaaa agctctgaag 4141 tcttcttcat ttaaattctt agatgatact tcatctggaa aattgtccca attagtagca 4201 tcacgctgtg agtaagttct aaaccatttt tttattgttg tattatctct aatcttacta 4261 ctcgatgagt tttcggtatt atctctattt ttaacttgga gcaggttcca ttcattgttt 4321 ttttcatcat agtgaataaa atcaactgct ttaacacttg tgcctgaaca ccatatccat 4381 ccggcgtaat acgactcact atagggagag cggccgccag atcttccgga tggctcgagt 4441 ttttcagcaa gatcttcgaa catcatcaat aatatacctt attttggatt gaagccaata 4501 tgataatgag ggggtggagt ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg 4561 tagtagtgtg gcggaagtgt gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg 4621 tggcaaaagt gacgtttttg gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg 4681 gttttaggcg gatgttgtag taaatttggg cgtaaccgag taagatttgg ccattttcgc 4741 gggaaaactg aataagagga agtgaaatct gaataatttt gtgttactca tagcgcgtaa 4801 tatttgtcta gggccgcggg gactttgacc gtttacgtgg agactcgccc aggtgttttt 4861 ctcaggtgtt ttccgcgttc cgggtcaaag ttggcgtttt attattatag tcagctgacg 4921 tgtagtgtat ttatacccgg gaattcggat ccaccggtga tatcttaatt aacggggtca 4981 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct 5041 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta 5101 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac 5161 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt 5221 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag 5281 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat 5341 gggcgtggat agcggtttga ctcacgggga tttccaagtc ttcaccccat tgacgtcaat 5401 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc 5461 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt 5521 ttagtgaacc gtcagatccc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg 5581 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg 5641 gcgagggcga gggcgatgcc //