LOCUS       Exported                4999 bp ds-DNA     circular SYN 24-SEP-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4999)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Sep 24, 2021 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4999
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             6..23
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     CDS             24..44
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /note="color: #cc99b2
                     Cleavage site after base 41"
                     /translation="ENLYFQG"
     misc_feature    46..51
                     /label=SD
                     /note="color: #a6acb3"
     CDS             60..62
                     /codon_start=1
                     /label=ATG
                     /note="color: #993366"
                     /translation="M"
     misc_feature    68..80
                     /label=DB
                     /note="color: #993366; direction: RIGHT"
     CDS             81..224
                     /codon_start=1
                     /label=GFP(s1->s3)
                     /note="color: #339966"
                     /translation="GEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFIC
                     TTG"
     misc_feature    186..188
                     /label=N40
                     /note="color: #00b2b2"
     misc_feature    225..317
                     /label=td intron
                     /note="This feature has 2 segments:
                        1: 225 .. 239 / #a6acb3
                        2: 240 .. 317 / #a6acb3"
     misc_feature    225..239
                     /label=P1
                     /note="color: #a6acb3"
     misc_feature    240..255
                     /label=P2
                     /note="color: #a6acb3"
     misc_feature    263..268
                     /label=P3
                     /note="color: #a6acb3"
     misc_feature    272..277
                     /label=P4
                     /note="color: #a6acb3"
     misc_feature    282..284
                     /label=P5
                     /note="color: #a6acb3"
     misc_feature    290..292
                     /label=P5'
                     /note="color: #a6acb3"
     misc_feature    296..301
                     /label=P4'
                     /note="color: #a6acb3"
     misc_feature    309..317
                     /label=P6a
                     /note="color: #a6acb3"
     terminator      324..352
                     /label=rrnC terminator
                     /note="color: #ffffff"
     terminator      376..474
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     terminator      388..474
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     misc_feature    550..1376
                     /label=p15A
                     /note="color: #ff2136"
     rep_origin      complement(815..1360)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
                     /note="color: #ffff00"
     promoter        1472..1500
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
                     /note="This forward directional feature has 3 segments:
                        1: 1472 .. 1477 / #ffffff / -35
                        2: 1478 .. 1494 / #ffffff
                        3: 1495 .. 1500 / #ffffff / -10"
     CDS             1548..2738
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /note="color: #ccffcc"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     misc_feature    2856..2863
                     /label=NotI
                     /note="color: #dff208"
     CDS             complement(3046..3924)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /note="color: #993366"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3951..4235
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
                     /note="color: #ffffff; direction: RIGHT"
     intron          4263..4419
                     /label=td intron
                     /note="color: #a6acb3"
     misc_feature    4268..4276
                     /label=P6a
                     /note="color: #a6acb3"
     misc_feature    4287..4293
                     /label=P7
                     /note="color: #a6acb3"
     misc_feature    4328..4333
                     /label=P3
                     /note="color: #a6acb3"
     misc_feature    4360..4365
                     /label=P7
                     /note="color: #a6acb3"
     misc_feature    4369..4375
                     /label=P9
                     /note="color: #a6acb3"
     misc_feature    4380..4386
                     /label=P9
                     /note="color: #a6acb3"
     misc_feature    4387..4391
                     /label=P9.1
                     /note="color: #a6acb3"
     misc_feature    4398..4402
                     /label=P9.1
                     /note="color: #a6acb3"
     misc_feature    4403..4408
                     /label=P9.2
                     /note="color: #a6acb3"
     misc_feature    4418..4419
                     /label=P9.2
                     /note="color: #a6acb3"
     intron          4420..4425
                     /label=3'H td intron
                     /note="color: #a6acb3"
     misc_feature    4420..4423
                     /label=P9.2
                     /note="color: #a6acb3"
     intron          4426..4434
                     /label=3'H td intron
                     /note="color: #a6acb3"
     CDS             4435..4437
                     /codon_start=1
                     /label=td 3'-exon
                     /note="color: #ff6600"
                     /translation="L"
     CDS             join(4438..4999,1..23)
                     /codon_start=1
                     /label=GFP(ih->end)
                     /note="color: #339966"
                     /translation="KLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQER
                     TISFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITAD
                     KQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEK
                     RDHMVLLEFVTAAGITHGMDELYKGSHHHHHH"
     misc_feature    4687..4689
                     /label=N136
                     /note="color: #00b2b2"
     gene            4729..4731
                     /label=N150
                     /note="color: #00b2b2; direction: RIGHT"
     misc_feature    4735..4737
                     /label=Y152
                     /note="color: #16de00"
     misc_feature    4843..4845
                     /label=P188
                     /note="color: #00b2b2"
ORIGIN
        1 gcagccacca tcatcatcac catgaaaacc tgtattttca gggcaaagga gatatacata
       61 tgcctagcat gaatcacaaa ggtgaagaac tgtttaccgg cgttgtgccg attctggtgg
      121 aactggatgg tgatgtgaat ggccataaat ttagcgttcg tggcgaaggc gaaggtgatg
      181 cgaccaacgg taaactgacc ctgaaattta tttgcaccac cggttaattg aggcctgagt
      241 ataaggtgac ttatacttgt aatctatcta aacggggaac ctctctagta gacaatcccg
      301 tgctaaattg taggactatg catatcctta gcgaaagcta aggatttttt ttgtctcccc
      361 atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
      421 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
      481 ggagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg
      541 gacatcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag
      601 tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga
      661 tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg
      721 cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt
      781 aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca
      841 agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat
      901 accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta
      961 ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc
     1021 gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc
     1081 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca
     1141 ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta
     1201 aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt
     1261 caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt
     1321 ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca
     1381 gataaaatat ttctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag
     1441 ccccatacga tataagttgt aattctcatg tttgacagct tatcatcgat aagctttaat
     1501 gcggtagttt atcacagtta aattgctaac gcagtcaggc accgtgtatg aaatctaaca
     1561 atgcgctcat cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata ggcttggtta
     1621 tgccggtact gccgggcctc ttgcgggata tcgtccattc cgacagcatc gccagtcact
     1681 atggcgtgct gctagcgcta tatgcgttga tgcaatttct atgcgcaccc gttctcggag
     1741 cactgtccga ccgctttggc cgccgcccag tcctgctcgc ttcgctactt ggagccacta
     1801 tcgactacgc gatcatggcg accacacccg tcctgtggat cctctacgcc ggacgcatcg
     1861 tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg
     1921 atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg
     1981 tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca ccattccttg
     2041 cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc
     2101 ataagggaga gcgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     2161 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg
     2221 taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg
     2281 cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct
     2341 tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg
     2401 cggccgacgc gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc
     2461 ccattatgat tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt
     2521 ccaggcaggt agatgacgac catcagggac agcttcaagg atcgctcgcg gctcttacca
     2581 gcctaacttc gatcattgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca
     2641 catggaacgg gttggcatgg attgtaggcg ccgccctata ccttgtctgc ctccccgcgt
     2701 tgcgtcgcgg tgcatggagc cgggccacct cgacctgaat ggaagccggc ggcacctcgc
     2761 taacggattc accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc
     2821 gcaaaccaac ccttggcaga acatatccat cgcgtgcggc cgcgtctccg ggagctgcat
     2881 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcagatca attcgcgcgc
     2941 gaaggcgaag cggcatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac
     3001 cctatgctac tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac
     3061 ggctacatca ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt
     3121 gcatttttta aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac
     3181 ggtggcgata ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc
     3241 ctcgcgccag cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga
     3301 cggcgacaag caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg
     3361 atcgctgatg tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc
     3421 gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc
     3481 cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg
     3541 cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt
     3601 aagccattca tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg
     3661 agcctccgga tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc
     3721 cggtcggcaa acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag
     3781 attgagaata taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt
     3841 ggcctcaatc ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag
     3901 gggatcattt tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa
     3961 ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac
     4021 caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac
     4081 aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca
     4141 cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg
     4201 acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc tacgctgaat
     4261 tcgaattggt tctacataaa tgcctaacga ctatcccttt ggggagtagg gtcaagtgac
     4321 tcgaaacgat agacaacttg ctttaacaag ttggagatat agtctgctct gcatggtgac
     4381 atgcagctgg atataattcc ggggtaagat taacgacctt atctgaacat aatgctaaaa
     4441 ctgccggttc cgtggccgac cctggtgacc accctgacct atggcgttca gtgctttagc
     4501 cgctatccgg atcatatgaa acgccatgat ttctttaaaa gcgcgatgcc ggaaggctat
     4561 gtgcaggaac gtaccattag cttcaaagat gatggcacct ataaaacccg tgcggaagtt
     4621 aaatttgaag gcgataccct ggtgaaccgc attgaactga aaggtattga ttttaaagaa
     4681 gatggcaaca ttctgggtca taaactggaa tataatttca acagccataa cgtgtatatt
     4741 accgccgata aacagaaaaa tggcatcaaa gcgaacttta aaatccgtca caacgtggaa
     4801 gatggtagcg tgcagctggc ggatcattat cagcagaata ccccgattgg tgatggcccg
     4861 gtgctgctgc cggataatca ttatctgagc acccagagcg ttctgagcaa agatccgaat
     4921 gaaaaacgtg atcatatggt gctgctggaa tttgttaccg ccgcgggcat tacccacggt
     4981 atggatgaac tgtataaag
//
