LOCUS       sequence-351749-        7845 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Block 6 with deletion in E3 and insertion of GFP expression 
            cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7845)
  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 7845)
  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-B6 ∆E3-GFP
FEATURES             Location/Qualifiers
     source          1..7845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     17..35
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     341..360
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        341..359
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        3267..3570
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3571..3774
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3725..3745
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3798..3807
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3804..3806,3807..3809,3810..4523)
                     /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(3804..3806,3807..3809,3810..4520)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3849..3870)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4110..4129)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4457..4478
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4614..4633)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4679..4734
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4733..4752)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5095..5111)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5095..5111)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    5119..5135
                     /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(5713..5719,5720..5737,5738..5743))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    5758..5779
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5899..5916)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6070..6658)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6150..6169)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6829..7620,7621..7689))
                     /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     7452..7471
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7690..7794)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattaacc tataaaaata ggcgtatcac gaggccgccc ctgcagccga attatattat
       61 ttttgccaaa taatttttaa caaaagctct gaagtcttct tcatttaaat tcttagatga
      121 tacttcatct ggaaaattgt cccaattagt agcatcacgc tgtgagtaag ttctaaacca
      181 tttttttatt gttgtattat ctctaatctt actactcgat gagttttcgg tattatctct
      241 atttttaact tggagcaggt tccattcatt gtttttttca tcatagtgaa taaaatcaac
      301 tgctttaaca cttgtgcctg aacaccatat ccatccggcg taatacgact cactataggg
      361 agagcggccg ccagatcttc cggatggctc gagtttttca gcaagatctt cgaagcgacg
      421 caaactaatg atggccgcag tgctcgttac cgtggagctt gagtgcatgc agcggttctt
      481 tgctgacccg gagatgcagc gcaagctaga ggaaacattg cactacacct ttcgacaggg
      541 ctacgtacgc caggcctgca agatctccaa cgtggagctc tgcaacctgg tctcctacct
      601 tggaattttg cacgaaaacc gccttgggca aaacgtgctt cattccacgc tcaagggcga
      661 ggcgcgccgc gactacgtcc gcgactgcgt ttacttattt ctatgctaca cctggcagac
      721 ggccatgggc gtttggcagc agtgcttgga ggagtgcaac ctcaaggagc tgcagaaact
      781 gctaaagcaa aacttgaagg acctatggac ggccttcaac gagcgctccg tggccgcgca
      841 cctggcggac atcattttcc ccgaacgcct gcttaaaacc ctgcaacagg gtctgccaga
      901 cttcaccagt caaagcatgt tgcagaactt taggaacttt atcctagagc gctcaggaat
      961 cttgcccgcc acctgctgtg cacttcctag cgactttgtg cccattaagt accgcgaatg
     1021 ccctccgccg ctttggggcc actgctacct tctgcagcta gccaactacc ttgcctacca
     1081 ctctgacata atggaagacg tgagcggtga cggtctactg gagtgtcact gtcgctgcaa
     1141 cctatgcacc ccgcaccgct ccctggtttg caattcgcag ctgcttaacg aaagtcaaat
     1201 tatcggtacc tttgagctgc agggtccctc gcctgacgaa aagtccgcgg ctccggggtt
     1261 gaaactcact ccggggctgt ggacgtcggc ttaccttcgc aaatttgtac ctgaggacta
     1321 ccacgcccac gagattaggt tctacgaaga ccaatcccgc ccgcctaatg cggagcttac
     1381 cgcctgcgtc attacccagg gccacattct tggccaattg caagccatca acaaagcccg
     1441 ccaagagttt ctgctacgaa agggacgggg ggtttacttg gacccccagt ccggcgagga
     1501 gctcaaccca atccccccgc cgccgcagcc ctatcagcag cagccgcggg cccttgcttc
     1561 ccaggatggc acccaaaaag aagctgcagc tgccgccgcc acccacggac gaggaggaat
     1621 actgggacag tcaggcagag gaggttttgg acgaggagga ggaggacatg atggaagact
     1681 gggagagcct agacgaggaa gcttccgagg tcgaagaggt gtcagacgaa acaccgtcac
     1741 cctcggtcgc attcccctcg ccggcgcccc agaaatcggc aaccggttcc agcatggcta
     1801 caacctccgc tcctcaggcg ccgccggcac tgcccgttcg ccgacccaac cgtagatggg
     1861 acaccactgg aaccagggcc ggtaagtcca agcagccgcc gccgttagcc caagagcaac
     1921 aacagcgcca aggctaccgc tcatggcgcg ggcacaagaa cgccatagtt gcttgcttgc
     1981 aagactgtgg gggcaacatc tccttcgccc gccgctttct tctctaccat cacggcgtgg
     2041 ccttcccccg taacatcctg cattactacc gtcatctcta cagcccatac tgcaccggcg
     2101 gcagcggcag caacagcagc ggccacacag aagcaaaggc gaccggatag caagactctg
     2161 acaaagccca agaaatccac agcggcggca gcagcaggag gaggagcgct gcgtctggcg
     2221 cccaacgaac ccgtatcgac ccgcgagctt agaaacagga tttttcccac tctgtatgct
     2281 atatttcaac agagcagggg ccaagaacaa gagctgaaaa taaaaaacag gtctctgcga
     2341 tccctcaccc gcagctgcct gtatcacaaa agcgaagatc agcttcggcg cacgctggaa
     2401 gacgcggagg ctctcttcag taaatactgc gcgctgactc ttaaggacta gtttcgcgcc
     2461 ctttctcaaa tttaagcgcg aaaactacgt catctccagc ggccacaccc ggcgccagca
     2521 cctgttgtca gcgccattat gagcaaggaa attcccacgc cctacatgtg gagttaccag
     2581 ccacaaatgg gacttgcggc tggagctgcc caagactact caacccgaat aaactacatg
     2641 agcgcgggac cccacatgat atcccgggtc aacggaatac gcgcccaccg aaaccgaatt
     2701 ctcctggaac aggcggctat taccaccaca cctcgtaata accttaatcc ccgtagttgg
     2761 cccgctgccc tggtgtacca ggaaagtccc gctcccacca ctgtggtact tcccagagac
     2821 gcccaggccg aagttcagat gactaactca ggggcgcagc ttgcgggcgg ctttcgtcac
     2881 agggtgcggt cgcccgggca gggtataact cacctgacaa tcagagggcg aggtattcag
     2941 ctcaacgacg agtcggtgag ctcctcgctt ggtctccgtc cggacgggac atttcagatc
     3001 ggcggcgccg gccgctcttc attcacgcct cgtcaggcaa tcctaactct gcagacctcg
     3061 tcctctgagc cgcgctctgg aggcattgga actctgcaat ttattgagga gtttgtgcca
     3121 tcggtctact ttaacccctt ctcgggacct cccggccact atccggatca atttattcct
     3181 aactttgacg cggtaaagga ctcggcggac ggctacgact gaaggatctc ggggtcatta
     3241 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     3301 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     3361 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     3421 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     3481 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     3541 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     3601 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     3661 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     3721 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta
     3781 gtgaaccgtc agatcccgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     3841 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     3901 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     3961 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     4021 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     4081 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     4141 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     4201 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     4261 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     4321 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     4381 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     4441 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     4501 gcatggacga gctgtacaag taaagcggcc gcactcctca ggtgcaggct gcctatcaga
     4561 aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat cgatcttttt
     4621 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     4681 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     4741 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     4801 gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag
     4861 tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt
     4921 tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc
     4981 ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc
     5041 cctcttctct tatggagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata
     5101 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccgggga
     5161 tccatatgtc gacggtctca aagatcttat tccctttaac taataaaaaa aaataataaa
     5221 gcatcactta cttaaaatca gttagcaaat ttctgtccag tttattcagc attcgaacat
     5281 ctttctagaa gatctcctac aatattctca gctgccatgg aaaatcgatg ttcttctttt
     5341 attctctcaa gattttcagg ctgtatatta aaacttatat taagaactat gctaaccacc
     5401 tcatcaggaa ccgttgtagg tggcgtgggt tttcttggca atcgactctc atgaaaacta
     5461 cgagctaaat attcaatatg ttcctcttga ccaactttat tctgcatttt ttttgaacga
     5521 ggtttagagc aagcttcagg aaactgagac aggaatttta ttaaaaattt aaattttgaa
     5581 gaaagttcag ggttaatagc atccattttt tgctttgcaa gttcctcagc attcttaaca
     5641 aaagacgtct cttttgacat gtttaaagtt taaacctcct gtgtgaaatt attatccgct
     5701 cataattcca cacattatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5761 agtgagctaa ctcacattaa ttgcgttgcg ctcactgcca attgctttcc agtcgggaaa
     5821 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5881 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5941 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6001 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6061 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6121 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6181 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6241 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6301 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6361 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6421 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6481 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6541 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6601 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6661 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6721 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6781 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6841 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6901 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6961 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7021 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7081 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7141 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7201 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7261 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7321 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7381 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7441 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7501 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7561 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7621 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7681 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7741 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7801 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcat
//
