LOCUS       CA-0234_CMVTO-sumIL2-re 5681 bp ds-DNA     circular     26-NOV-2024
DEFINITION  .
COMMENT     Concatenation of 4 sequences.
FEATURES             Location/Qualifiers
     CDS             complement(570..1430)
                     /label="Amp (bla)"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     CDS             complement(573..1232)
                     /label="AmpR"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     promoter        complement(1431..1529)
                     /label="bla promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     promoter        1793..2420
                     /label="CMV"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     promoter        1872..2175
                     /label="CMV enhancer"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     promoter        2176..2379
                     /label="CMV promoter"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          2330..2350
                     /label="CMV-Forward Genewiz"
                     /note="sequence: CGCAAATGGGCGGTAGGCGTG"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     TATA_signal     2365..2371
                     /label="TATA box"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    2381..2399
                     /label="TetO"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    2381..2420
                     /label="TetR binding sites"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    2402..2420
                     /label="TetO"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          2481..2501
                     /label="CMVF1"
                     /note="sequence: tgacctccatagaagacaccg"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          2502..2517
                     /label="ToFlpF"
                     /note="sequence: GGACCGATCCAGCCTC"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_feature    2527..2543
                     /label="Inf FWD"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          2527..2548
                     /label="CA-0084.F"
                     /note="sequence: gcgtttaaacttaagcttcgcc"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer          2527..2563
                     /label="IL2.F.1"
                     /note="sequence: gcgtttaaacttaagcttcgccaccATGTACAGGATG"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    2544..2551
                     /label="Kozak"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    2552..2614
                     /label="IL-2 Signal Peptide"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     CDS             2552..2614
                     /label="Translation 2552-2614"
                     /translation="MYRMQLLSCIALSLALVTNAS"
     misc_feature    2615..3067
                     /label="SumIL2"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     CDS             2615..3067
                     /label="Translation 2615-3067"
                     /translation="APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTAKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNYIHPFHLVIHNESGGGGSFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLT"
     misc_feature    2821..2845
                     /label="PCR REV"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          complement(2821..2870)
                     /label="CA-0161.R"
                     /note="sequence: TAACCAGATGGAATGGGTGAATATAATTTTTGCTTTGGGCGAGATTAAGC"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    2846..2884
                     /label="Renin cleavage site"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    2855..2870
                     /label="INF"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          2855..2918
                     /label="CA-0161.F"
                     /note="sequence: CCATTCCATCTGGTTATTCATAACGAGTCGGGCGGTGGGGGATCCTTCCACTTTGATCCGCGGG"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    2885..2899
                     /label="linker"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    2900..2918
                     /label="PCR FWD"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          complement(3035..3079)
                     /label="CA-0025.R"
                     /note="sequence: gTTCgggccCTAGGTCAAAGTAGAAATTATACTCTGGCAAAATGT"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    3067..3079
                     /label="Inf REV"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     CDS             3068..3070
                     /label="Translation 3068-3070"
                     /translation="*"
     primer          complement(3070..3091)
                     /label="CA-0127.R2"
                     /note="sequence: agatgagtttttgTTCgggccC"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     CDS             3079..3108
                     /label="c-myc epitope"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     misc_feature    3108..3110
                     /label="New Stop"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     CDS             3121..3138
                     /label="Polyhistidine tag"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     misc_feature    3139..3141
                     /label="STOP (missing now)"
                     /ApEinfo_revcolor="#a187be"
                     /ApEinfo_fwdcolor="#a187be"
     polyA_signal    3160..3374
                     /label="BGH polyadenylation sequence"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          complement(3161..3178)
                     /label="BGHR Genewiz"
                     /note="sequence: TAGAAGGCACAGTCGAGG"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     polyA_signal    3407..3430
                     /label="BGH pA"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    3713..3760
                     /label="FRT"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_recomb     3727..3760
                     /label="FRT"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     gene            3768..4788
                     /label="Hygromycin"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     polyA_signal    4920..5050
                     /label="SV40 pA"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer_bind     complement(5086..5106)
                     /label="M13-rev"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_binding    complement(5112..5134)
                     /label="LacO"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     promoter        complement(5138..5168)
                     /label="lac promoter"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     rep_origin      5433..425
                     /label="pUC ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     rep_origin      complement(5474..475)
                     /label="ColE1 origin"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
ORIGIN
        1 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
       61 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      121 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      181 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      241 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
      301 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
      361 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
      421 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
      481 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
      541 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
      601 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
      661 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
      721 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
      781 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
      841 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
      901 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
      961 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1021 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1081 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     1141 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     1201 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     1261 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     1321 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     1381 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     1441 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     1501 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     1561 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     1621 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     1681 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     1741 atctgcttag ggttaggcgt tttgcgctgc ttcgtacggg ccagatatac gcgttgacat
     1801 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     1861 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     1921 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     1981 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     2041 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     2101 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     2161 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     2221 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     2281 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     2341 ggtaggcgtg tacggtggga ggtctatata agcagagctc tccctatcag tgatagagat
     2401 ctccctatca gtgatagaga tcgtcgacga gctcgtttag tgaaccgtca gatcgcctgg
     2461 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
     2521 actctagcgt ttaaacttaa gcttcgccac cATGTACAGG ATGCAACTCC TGTCTTGCAT
     2581 TGCACTAAGT CTTGCACTTG TCACAAACgc tAGCGCTCCA ACCTCCAGCT CTACAAAAAA
     2641 GACTCAGCTG CAGCTGGAGC ACCTGCTGCT CGACCTACAA ATGATACTCA ACGGAATCAA
     2701 CAACTATAAG AATCCTAAGC TGACGCGGAT GCTGACCGCC AAGTTTTACA TGCCCAAAAA
     2761 GGCGACCGAG CTGAAGCATT TGCAGTGTCT GGAGGAAGAG CTCAAGCCTT TGGAGGAGGT
     2821 GCTTAATCTC GCCCAAAGCA AAAATTATAT TCACCCATTC CATCTGGTTA TTCATAACGA
     2881 GTCGGGCGGT GGGGGATCCT TCCACTTTGA TCCGCGGGAC GTCGTGTCAA ATATCAACGT
     2941 CTTTGTGCTG GAACTGAAAG GCTCTGAGAC AACCTTTATG TGTGAATATG CCGATGAAAC
     3001 TGCTACCATC GTTGAGTTTT TAAATCGATG GATAACATTT TGCCAGAGTA TAATTTCTAC
     3061 TTTGACCTAG ggcccGAAca aaaactcatc tcagaagagg atctgaatag cgccgtcgac
     3121 catcatcata atcatcattg agtttaaacc cgctgatcag cctcgactgt gccttctagt
     3181 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3241 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3301 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3361 aggcatgctg gggatgcggt gggctctatg gagatctatc gatgttctgg ggatgcggtg
     3421 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3481 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3541 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3601 gccggctttc cccgtcaagc tctaaatcgg gggtcccttt agggttccga tttagtgctt
     3661 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtacc tagaagttcc
     3721 tattccgaag ttcctattct ctagaaagta taggaacttc cttggccaaa aagcctgaac
     3781 tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga
     3841 tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat
     3901 atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc
     3961 actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga
     4021 gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa
     4081 ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg
     4141 atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta
     4201 catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga
     4261 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg
     4321 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga
     4381 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc
     4441 aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga
     4501 cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata
     4561 tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg
     4621 cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc
     4681 gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg
     4741 ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag cacgtactac
     4801 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4861 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4921 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4981 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5041 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5101 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5161 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5221 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5281 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5341 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5401 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5461 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5521 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5581 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5641 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct t
//