LOCUS       CA-0061_CMVTO-SumIL2-2G 6353 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    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"
     CDS             2615..3001
                     /label="Translation 2615-3001"
                     /translation="APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTAKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSII"
     misc_feature    2615..3013
                     /label="SumIL2"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    3002..3016
                     /label="FWD"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer          3002..3092
                     /label="CA-0005.F"
                     /note="sequence: TCTACTTTGACCGGTGGAGGGGGTTCTGGCGGTGGGGGATCTTATATTCACCCATTCCATCTGGTTATTCATAACGAGTCGGGCGGTGGGG"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     CDS             3002..3097
                     /label="Translation 3002-3097"
                     /translation="STLTGGGGSGGGGSYIHPFHLVIHNESGGGGS"
     primer          complement(3017..3107)
                     /label="CA-0005.R"
                     /note="sequence: CATTCACCGCGGATCCCCCACCGCCCGACTCGTTATGAATAACCAGATGGAATGGGTGAATATAAGATCCCCCACCGCCAGAACCCCCTCC"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    3044..3082
                     /label="Renin cleavage site"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    3093..3107
                     /label="Rev"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     CDS             3098..3107
                     /label="Translation 3098-3107"
                     /translation="AVN"
     misc_feature    3098..3739
                     /label="hIL-2Rbeta ectodomain"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     CDS             3107..3739
                     /label="Translation 3107-3739"
                     /translation="GTSQFTCFYNSRANISCVWSQDGALQDTSCQVHAWPDRRRWNQTCELLPVSQASWACNLILGAPDSQKLTTVDIVTLRVLCREGVRWRVMAIQDFKPFENLRLMAPISLQVVHVETHRCNISWEISQASHYFERHLEFEARTLSPGHTWEEAPLLTLKQKQEWICLETLTPDTQYEFQVRVKPLQGEFTTWSPWSQPLAFRTKPAALGKDT"
     primer          complement(3722..3765)
                     /label="IL2Rb.REV.1"
                     /note="sequence: tgagatgagtttttgTTCgggccCTAGGTGTCCTTCCCAAGGGC"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     CDS             3740..3742
                     /label="Translation 3740-3742"
                     /translation="*"
     CDS             3751..3780
                     /label="c-myc epitope"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     misc_feature    3780..3782
                     /label="New Stop"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     CDS             3793..3810
                     /label="Polyhistidine tag"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     misc_feature    3811..3813
                     /label="STOP (missing now)"
                     /ApEinfo_revcolor="#a187be"
                     /ApEinfo_fwdcolor="#a187be"
     polyA_signal    3832..4046
                     /label="BGH polyadenylation sequence"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          complement(3833..3850)
                     /label="BGHR Genewiz"
                     /note="sequence: TAGAAGGCACAGTCGAGG"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     polyA_signal    4079..4102
                     /label="BGH pA"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    4385..4432
                     /label="FRT"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_recomb     4399..4432
                     /label="FRT"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     gene            4440..5460
                     /label="Hygromycin"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     polyA_signal    5592..5722
                     /label="SV40 pA"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer_bind     complement(5758..5778)
                     /label="M13-rev"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_binding    complement(5784..5806)
                     /label="LacO"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     promoter        complement(5810..5840)
                     /label="lac promoter"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     rep_origin      6105..425
                     /label="pUC ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     rep_origin      complement(6146..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 AAAATTTCCA CTTTGATCCG CGGGACGTCG TGTCAAATAT
     2881 CAACGTCTTT GTGCTGGAAC TGAAAGGCTC TGAGACAACC TTTATGTGTG AATATGCCGA
     2941 TGAAACTGCT ACCATCGTTG AGTTTTTAAA TCGATGGATA ACATTTTGCC AGAGTATAAT
     3001 TTCTACTTTG ACCGGTGGAG GGGGTTCTGG CGGTGGGGGA TCTTATATTC ACCCATTCCA
     3061 TCTGGTTATT CATAACGAGT CGGGCGGTGG GGGATCCGCG GTGAATGGCA CTTCCCAGTT
     3121 CACATGCTTC TACAATTCGA GAGCCAACAT CTCCTGTGTC TGGAGCCAAG ATGGGGCTCT
     3181 GCAGGACACT TCCTGCCAAG TCCATGCCTG GCCGGACAGA CGGCGGTGGA ACCAAACCTG
     3241 TGAGCTGCTC CCCGTGAGTC AAGCATCCTG GGCCTGCAAC CTGATCCTCG GAGCCCCAGA
     3301 TTCTCAGAAA CTGACCACAG TTGACATCGT CACCCTGAGG GTGCTGTGCC GTGAGGGGGT
     3361 GCGATGGAGG GTGATGGCCA TCCAGGACTT CAAGCCCTTT GAGAACCTTC GCCTGATGGC
     3421 CCCCATCTCC CTCCAAGTTG TCCACGTGGA GACCCACAGA TGCAACATAA GCTGGGAAAT
     3481 CTCCCAAGCC TCCCACTACT TTGAAAGACA CCTGGAGTTC GAGGCCCGGA CGCTGTCCCC
     3541 AGGCCACACC TGGGAGGAGG CCCCCCTGCT GACTCTCAAG CAGAAGCAGG AATGGATCTG
     3601 CCTGGAGACG CTCACCCCAG ACACCCAGTA TGAGTTTCAG GTGCGGGTCA AGCCTCTGCA
     3661 AGGCGAGTTC ACGACCTGGA GCCCCTGGAG CCAGCCCCTG GCCTTCAGGA CAAAGCCTGC
     3721 AGCCCTTGGG AAGGACACCT AGggcccGAA caaaaactca tctcagaaga ggatctgaat
     3781 agcgccgtcg accatcatca taatcatcat tgagtttaaa cccgctgatc agcctcgact
     3841 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3901 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3961 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     4021 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggagatcta tcgatgttct
     4081 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     4141 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4201 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4261 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggtccct ttagggttcc
     4321 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4381 cctagaagtt cctattccga agttcctatt ctctagaaag tataggaact tccttggcca
     4441 aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg
     4501 tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag
     4561 gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt
     4621 atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg
     4681 aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag
     4741 acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga
     4801 tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg
     4861 gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact
     4921 ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga
     4981 tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca
     5041 acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt
     5101 tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta
     5161 tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc
     5221 tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca
     5281 atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg
     5341 ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg
     5401 tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat
     5461 agcacgtact acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5521 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5581 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5641 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5701 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5761 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5821 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5881 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5941 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6001 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6061 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6121 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6181 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6241 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6301 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctt
//