LOCUS       CA-0013_CMVTO-SumIL2-re 6338 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"
     misc_feature    2615..3013
                     /label="SumIL2"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     CDS             2615..3082
                     /label="Translation 2615-3082"
                     /translation="APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTAKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLTGAGTGYIHPFHLVIHNESGGGGS"
     misc_feature    3014..3028
                     /label="PCR REV"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    3029..3067
                     /label="Renin cleavage site"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    3068..3082
                     /label="PCR FWD"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_feature    3083..3724
                     /label="hIL-2Rbeta ectodomain"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     CDS             3083..3724
                     /label="Translation 3083-3724"
                     /translation="AVNGTSQFTCFYNSRANISCVWSQDGALQDTSCQVHAWPDRRRWNQTCELLPVSQASWACNLILGAPDSQKLTTVDIVTLRVLCREGVRWRVMAIQDFKPFENLRLMAPISLQVVHVETHRCNISWEISQASHYFERHLEFEARTLSPGHTWEEAPLLTLKQKQEWICLETLTPDTQYEFQVRVKPLQGEFTTWSPWSQPLAFRTKPAALGKDT"
     primer          complement(3707..3750)
                     /label="IL2Rb.REV.1"
                     /note="sequence: tgagatgagtttttgTTCgggccCTAGGTGTCCTTCCCAAGGGC"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     CDS             3725..3727
                     /label="Translation 3725-3727"
                     /translation="*"
     CDS             3736..3765
                     /label="c-myc epitope"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     misc_feature    3765..3767
                     /label="New Stop"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     CDS             3778..3795
                     /label="Polyhistidine tag"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     misc_feature    3796..3798
                     /label="STOP (missing now)"
                     /ApEinfo_revcolor="#a187be"
                     /ApEinfo_fwdcolor="#a187be"
     polyA_signal    3817..4031
                     /label="BGH polyadenylation sequence"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          complement(3818..3835)
                     /label="BGHR Genewiz"
                     /note="sequence: TAGAAGGCACAGTCGAGG"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     polyA_signal    4064..4087
                     /label="BGH pA"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    4370..4417
                     /label="FRT"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_recomb     4384..4417
                     /label="FRT"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     gene            4425..5445
                     /label="Hygromycin"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     polyA_signal    5577..5707
                     /label="SV40 pA"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer_bind     complement(5743..5763)
                     /label="M13-rev"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_binding    complement(5769..5791)
                     /label="LacO"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     promoter        complement(5795..5825)
                     /label="lac promoter"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     rep_origin      6090..425
                     /label="pUC ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     rep_origin      complement(6131..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 ACCggcgccg ggacaggaTA TATTCACCCA TTCCATCTGG TTATTCATAA
     3061 CGAGTCGGGC GGTGGGGGAT CCGCGGTGAA TGGCACTTCC CAGTTCACAT GCTTCTACAA
     3121 TTCGAGAGCC AACATCTCCT GTGTCTGGAG CCAAGATGGG GCTCTGCAGG ACACTTCCTG
     3181 CCAAGTCCAT GCCTGGCCGG ACAGACGGCG GTGGAACCAA ACCTGTGAGC TGCTCCCCGT
     3241 GAGTCAAGCA TCCTGGGCCT GCAACCTGAT CCTCGGAGCC CCAGATTCTC AGAAACTGAC
     3301 CACAGTTGAC ATCGTCACCC TGAGGGTGCT GTGCCGTGAG GGGGTGCGAT GGAGGGTGAT
     3361 GGCCATCCAG GACTTCAAGC CCTTTGAGAA CCTTCGCCTG ATGGCCCCCA TCTCCCTCCA
     3421 AGTTGTCCAC GTGGAGACCC ACAGATGCAA CATAAGCTGG GAAATCTCCC AAGCCTCCCA
     3481 CTACTTTGAA AGACACCTGG AGTTCGAGGC CCGGACGCTG TCCCCAGGCC ACACCTGGGA
     3541 GGAGGCCCCC CTGCTGACTC TCAAGCAGAA GCAGGAATGG ATCTGCCTGG AGACGCTCAC
     3601 CCCAGACACC CAGTATGAGT TTCAGGTGCG GGTCAAGCCT CTGCAAGGCG AGTTCACGAC
     3661 CTGGAGCCCC TGGAGCCAGC CCCTGGCCTT CAGGACAAAG CCTGCAGCCC TTGGGAAGGA
     3721 CACCTAGggc ccGAAcaaaa actcatctca gaagaggatc tgaatagcgc cgtcgaccat
     3781 catcataatc atcattgagt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     3841 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3901 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3961 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     4021 catgctgggg atgcggtggg ctctatggag atctatcgat gttctgggga tgcggtgggc
     4081 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     4141 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4201 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4261 ggctttcccc gtcaagctct aaatcggggg tccctttagg gttccgattt agtgctttac
     4321 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtacctag aagttcctat
     4381 tccgaagttc ctattctcta gaaagtatag gaacttcctt ggccaaaaag cctgaactca
     4441 ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc
     4501 agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg
     4561 tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact
     4621 ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc agcgagagcc
     4681 tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg
     4741 aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc
     4801 ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat
     4861 ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg
     4921 acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg
     4981 actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg
     5041 acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat
     5101 acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc
     5161 gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc
     5221 tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag
     5281 cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta
     5341 cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg
     5401 atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac gtactacgag
     5461 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5521 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     5581 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5641 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5701 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5761 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5821 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5881 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5941 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6001 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6061 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6121 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6181 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6241 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6301 acctgtccgc ctttctccct tcgggaagcg tggcgctt
//