LOCUS       sequence-351752-        5660 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Block 1 with deletion in E1 and insertion of GFP expression 
            cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5660)
  AUTHORS   Jang Y, Bunz F
  TITLE     AdenoBuilder: A platform for the modular assembly of recombinant 
            adenoviruses.
  JOURNAL   STAR Protoc. 2022 Jan 20;3(1):101123. doi: 
            10.1016/j.xpro.2022.101123. eCollection 2022 Mar 18.
  PUBMED    35098167
REFERENCE   2  (bases 1 to 5660)
  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: "STAR 
            Protoc."; date: "2022-01-20"; volume: "3(1):101123. doi"; pages: " 
            10.1016/j.xpro.2022.101123. eCollection 2022 Mar 18"
COMMENT     Sequence Label: pAd5-B1-deltaE1-GFP
FEATURES             Location/Qualifiers
     source          1..5660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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(join(1914..1920,1921..1938,1939..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(join(3030..3821,3822..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
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5547..5549,5550..5552,5553..5660,1..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             join(5547..5549,5550..5552,5553..5660,1..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
//
