LOCUS       Exported                4906 bp ds-DNA     circular SYN 01-JUN-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4906)
  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..4906
                     /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"
     enhancer        319..622
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        623..826
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     777..797
                     /label=CMV-Forward
                     /note="Genewiz primer collection"
     primer_bind     830..850
                     /label=JD133
                     /note="fp1; template pJD78(2) e2a gene in pJD07; dig pcr 
                     amp product with PacI, NotI; backbone pJD07 dig with PacI, 
                     NotI"
     primer_bind     831..854
                     /label=JD132
                     /note="use for CMV-E2A cloning; use instead of JD115 for 
                     pJD78(2) construct as template"
     misc_feature    837..943
                     /label=e2A 5utr (intron removed)
                     /note="Ad2"
     CDS             complement(837..857)
                     /codon_start=1
                     /product="33K"
                     /label=33K
                     /protein_id="AP_000179.1"
                     /translation="RSCLYH"
     misc_feature    858..934
                     /label=exon
     CDS             complement(858..934)
                     /codon_start=1
                     /product="100K"
                     /label=100K
                     /protein_id="AP_000178.1"
                     /translation="DIASLDEVPKIFEGLGRDEKRAANA"
     misc_feature    950..959
                     /label=kozak
     misc_feature    956..2562
                     /label=Ad2 bases 22403-24079
     CDS             956..2545
                     /codon_start=1
                     /product="DBP"
                     
                     /protein_id="AP_000177.1"
                     /translation="MASREEEQRETTPERGRGAARRPPTMEDVSSPSPSPPPPRAPPKK
                     RLRRRLESEDEEDSSQDALVPRTPSPRPSTSTADLAIASKKKKKRPSPKPERPPSPEVI
                     VDSEEEREDVALQMVGFSNPPVLIKHGKGGKRTVRRLNEDDPVARGMRTQEEKEESSEA
                     ESESTVINPLSLPIVSAWEKGMEAARALMDKYHVDNDLKANFKLLPDQVEALAAVCKTW
                     LNEEHRGLQLTFTSNKTFVTMMGRFLQAYLQSFAEVTYKHHEPTGCALWLHRCAEIEGE
                     LKCLHGSIMINKEHVIEMDVTSENGQRALKEQSSKAKIVKNRWGRNVVQISNTDARCCV
                     HDAACPANQFSGKSCGMFFSEGAKAQVAFKQIKAFMQALYPNAQTGHGHLLMPLRCECN
                     SKPGHAPFLGRQLPKLTPFALSNAEDLDADLISDKSVLASVHHPALIVFQCCNPVYRNS
                     RAQGGGPNCDFKISAPDLLNALVMVRSLWSENFTELPRMVVPEFKWSTKHQYRNVSLPV
                     AHSDARQNPFDF"
     misc_feature    1083..1086
                     /label=overhang2 donor plasmid
     misc_feature    1105
                     /label=remove BsmBI site
                     /note="/silent base substitutionT to G"
     misc_feature    1126
                     /label=remove BbsI site
                     /note="/A to G silent base substitution"
     misc_feature    1384
                     /label=remove BbsI site
                     /note="/A to G silent base substitution"
     primer_bind     1430..1449
                     /label=JD90
                     /note="e2a gene sequencing primer"
     primer_bind     complement(1541..1563)
                     /label=JD91
                     /note="e2a gene sequencing primer"
     primer_bind     complement(2542..2567)
                     /label=JD116
     misc_feature    2546..2562
                     /label=3UTR Ad2E2A
                     /note="/NCBI AC_000007"
     misc_feature    2563..2571
                     /label=spacer
     misc_feature    2572..2581
                     /label=AT rich spacer
     polyA_signal    2582..2806
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    2672..2677
                     /label=signal
     misc_feature    2815..2857
                     /label=downstream MCS
                     /note="downstream multiple cloning site"
     primer_bind     complement(2862..2887)
                     /label=Reverse (CCW) Analysis Primer
     primer_bind     complement(2868..2887)
                     /label=T7
                     /note="Genewiz primer collection"
     promoter        complement(2869..2887)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2979..3083
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3084..3944
                     /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"
     misc_feature    3945..4028
                     /label=terminator sequence?
     primer_bind     complement(4016..4034)
                     /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(4029..4041)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4106..4124)
                     /label=PR-JD32
                     /note="pJD02/03 sequencing primer"
     rep_origin      4142..4730
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4666..4685
                     /label=PR-JD29
                     /note="pJD11 rtTA CDS, pA sequencing primer"
     primer_bind     4840..4858
                     /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 ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      361 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      421 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      481 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      541 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      601 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      661 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      721 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      781 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcaga cctaggttgt
      841 gatacaggca gctgcggagc gtttgccgcg cgtttctcgt cgcgtccaag accctcaaag
      901 atttttggca cttcgtcgag cgaggcgata tcaggagaag gaaggtaccg ccagcatggc
      961 cagtcgggaa gaggagcagc gcgaaaccac ccccgagcgc ggacgcggtg cggcgcgacg
     1021 tccaccaacc atggaggacg tgtcgtcccc gtcgccgtcg ccgccgcctc cccgcgcgcc
     1081 cccaaaaaag cggctgaggc ggcggctcga gtccgaggac gaagaggact cgtcacaaga
     1141 tgcgctggtg ccgcgcacac ccagcccgcg gccatcgacc tcgacggcgg atttggccat
     1201 tgcgtccaaa aagaaaaaga agcgcccctc tcccaagccc gagcgcccgc catccccaga
     1261 ggtgatcgtg gacagcgagg aagaaagaga agatgtggcg ctacaaatgg tgggtttcag
     1321 caacccaccg gtgctaatca agcacggcaa gggaggtaag cgcacggtgc ggcggctgaa
     1381 tgaggacgac ccagtggcgc ggggtatgcg gacgcaagag gaaaaggaag agtccagtga
     1441 agcggaaagt gaaagcacgg tgataaaccc gctgagcctg ccgatcgtgt ctgcgtggga
     1501 gaagggcatg gaggctgcgc gcgcgttgat ggacaagtac cacgtggata acgatctaaa
     1561 ggcaaacttc aagctactgc ctgaccaagt ggaagctctg gcggccgtat gcaagacctg
     1621 gctaaacgag gagcaccgcg ggttgcagct gaccttcacc agcaacaaga cctttgtgac
     1681 gatgatgggg cgattcctgc aggcgtacct gcagtcgttt gcagaggtaa cctacaagca
     1741 ccacgagccc acgggctgcg cgttgtggct gcaccgctgc gctgagatcg aaggcgagct
     1801 taagtgtcta cacgggagca ttatgataaa taaggagcac gtgattgaaa tggatgtgac
     1861 gagcgaaaac gggcagcgcg cgctgaagga gcagtctagc aaggccaaga tcgtgaagaa
     1921 ccggtggggc cgaaatgtgg tgcagatctc caacaccgac gcaaggtgct gcgtgcatga
     1981 cgcggcctgt ccggccaatc agttttccgg caagtcttgc ggcatgttct tctctgaagg
     2041 cgcaaaggct caggtggctt ttaagcagat caaggctttc atgcaggcgc tgtatcctaa
     2101 cgcccagacc gggcacggtc accttctgat gccactacgg tgcgagtgca actcaaagcc
     2161 tgggcatgca ccctttttgg gaaggcagct accaaagttg actccgttcg ccctgagcaa
     2221 cgcggaggac ctggacgcgg atctgatctc cgacaagagc gtgctggcca gcgtgcacca
     2281 cccggcgctg atagtgttcc agtgctgcaa ccctgtgtat cgcaactcgc gcgcgcaggg
     2341 cggaggcccc aactgcgact tcaagatatc ggcgcccgac ctgctaaacg cgttggtgat
     2401 ggtgcgcagc ctgtggagtg aaaacttcac cgagctgccg cggatggttg tgcctgagtt
     2461 taagtggagc actaaacacc agtatcgcaa cgtgtccctg ccagtggcgc atagcgatgc
     2521 gcggcagaac ccctttgatt tttaaacggc gcagacggca agatgccatt tatctatttt
     2581 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2641 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2701 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2761 tgggaacaca atagcaggca tgctggggat gcggtgggct ctatggccat tcctgtagac
     2821 ttcttaatta agacgtcaga attctcgagg cggccgcatg tgagtctccc tatagtgagt
     2881 cgtattaatt tcgcgggcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2941 tatccgctca tgagtagcac caggcgttta agggcacccg cggaacccct atttgtttat
     3001 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3061 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3121 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3181 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3241 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3301 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3361 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3421 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3481 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3541 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3601 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     3661 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3721 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3781 ctggagccgg tgagcgtgga tctcgcggta tcattgcagc actggggcca gatggtaagc
     3841 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3901 gacagatcgc tgagataggt gcgtcactga ttaagcattg gtaactgtca gaccaagttt
     3961 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4021 agatccttta cgtgccgatc aaagtctcat tttcgccaaa agttgtcatg accaaaatcc
     4081 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4141 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4201 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4261 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4321 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4381 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4441 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4501 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4561 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4621 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4681 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4741 atgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4801 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4861 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagc
//
