LOCUS       CA-0212_CMVTO-hIL2-rens 5642 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"
     primer          1525..1541
                     /label="AmpR_seq_F"
                     /note="sequence: ggggttccgcgcacatt"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     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          2527..2548
                     /label="CA-0084.F"
                     /note="sequence: gcgtttaaacttaagcttcgcc"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     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..3028
                     /label="Translation 2552-3028"
                     /translation="MYRMQLLSCIALSLALVTNASAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAYIHPFHLVIHNESPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLT"
     misc_feature    2615..3028
                     /label="Human IL-2"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          complement(2809..2858)
                     /label="CA-0137.R"
                     /note="sequence: TAACCAGATGGAATGGGTGAATATAAGCTAAATTTAGCACTTCCTCCAGA"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     misc_feature    2834..2872
                     /label="Renin cleavage site"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    2843..2858
                     /label="INF"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          2843..2897
                     /label="CA-0137.F"
                     /note="sequence: CCATTCCATCTGGTTATTCATAACGAGTCGCCCAGGGACTTAATCAGCAATATCA"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     CDS             3029..3031
                     /label="Translation 3029-3031"
                     /translation="*"
     primer          complement(3031..3052)
                     /label="CA-0127.R2"
                     /note="sequence: agatgagtttttgTTCgggccC"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     CDS             3040..3069
                     /label="c-myc epitope"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     misc_feature    3069..3071
                     /label="New Stop"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     CDS             3082..3099
                     /label="Polyhistidine tag"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     misc_feature    3100..3102
                     /label="STOP (missing now)"
                     /ApEinfo_revcolor="#a187be"
                     /ApEinfo_fwdcolor="#a187be"
     polyA_signal    3121..3335
                     /label="BGH polyadenylation sequence"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          complement(3122..3139)
                     /label="BGHR Genewiz"
                     /note="sequence: TAGAAGGCACAGTCGAGG"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     polyA_signal    3368..3391
                     /label="BGH pA"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    3674..3721
                     /label="FRT"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_recomb     3688..3721
                     /label="FRT"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     gene            3729..4749
                     /label="Hygromycin"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     polyA_signal    4881..5011
                     /label="SV40 pA"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer_bind     complement(5047..5067)
                     /label="M13-rev"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_binding    complement(5073..5095)
                     /label="LacO"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     promoter        complement(5099..5129)
                     /label="lac promoter"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     rep_origin      5394..425
                     /label="pUC ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     rep_origin      complement(5435..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 tAGCGCACCT ACTTCAAGTT CTACAAAGAA
     2641 AACACAGCTA CAACTGGAGC ATTTACTGCT GGATTTACAG ATGATTTTGA ATGGAATTAA
     2701 TAATTACAAG AATCCCAAAC TCACCAGGAT GCTCACATTT AAGTTTTACA TGCCCAAGAA
     2761 GGCCACAGAA CTGAAACATC TTCAGTGTCT AGAAGAAGAA CTCAAACCTC TGGAGGAAGT
     2821 GCTAAATTTA GCTTATATTC ACCCATTCCA TCTGGTTATT CATAACGAGT CGCCCAGGGA
     2881 CTTAATCAGC AATATCAACG TAATAGTTCT GGAACTAAAG GGATCTGAAA CAACATTCAT
     2941 GTGTGAATAT GCTGATGAGA CAGCAACCAT TGTAGAATTT CTGAACAGAT GGATTACCTT
     3001 TTGTCAAAGC ATCATCTCAA CACTGACTTA GggcccGAAc aaaaactcat ctcagaagag
     3061 gatctgaata gcgccgtcga ccatcatcat aatcatcatt gagtttaaac ccgctgatca
     3121 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3181 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3241 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3301 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggagatctat
     3361 cgatgttctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3421 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3481 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3541 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggtccctt
     3601 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3661 gttcacgtac ctagaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt
     3721 ccttggccaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     3781 tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct
     3841 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     3901 aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg
     3961 acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     4021 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca
     4081 tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
     4141 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
     4201 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
     4261 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
     4321 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
     4381 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
     4441 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat
     4501 cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
     4561 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
     4621 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg
     4681 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
     4741 caaaggaata gcacgtacta cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4801 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4861 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     4921 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4981 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     5041 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5101 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5161 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5221 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5281 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5341 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5401 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5461 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5521 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5581 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5641 tt
//