LOCUS       62056_21845        5376 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5376)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1446..1502
                     /codon_start=1
                     /label=GCN4_v4
                     /note="GCN4 peptide optimized to improve solubility while 
                     preserving binding to an scFv-GFP fusion protein (Tanenbaum
                     et al., 2014)"
                     /translation="EELLSKNYHLENEVARLKK"
     primer_bind     complement(3200..3216)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3224..3240)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3248..3278)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3293..3314)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3602..4190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4364..5176)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRFAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(5177..5281)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagactgtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gacgcgtatt gggatcctga
      421 agatcaaggc tgctgcagtg agctgtgatc gcactactgc actccagcct gggccacaga
      481 gtgaaccctg tctctaaaaa aaaataaaaa caaaacaaga gtccctgggc cctttgaata
      541 aaaagagagt aaagaggctg ggcacagtgg ctcacgcccg taatcccagc actttgggag
      601 gctgaggcag gcagatcact tgtgcccagg agtttgaggc cagcgtgggc aacatagcaa
      661 aatcccatct ctccaaaaaa aaaaaaaaaa ggaaaagaaa agaaagtgag taaaggcttt
      721 tgggttataa agagaaaggg agaaggatga tccagactag aaagaagatt attcctttct
      781 tagctcaacc tttttgctcc aaaaaaaaga ggaggaaggg gccaaacata gattatcctg
      841 caccagaggg ccagctcctt aaatctgagt ctcatcccaa ccaccatcat ttgtaaaaac
      901 caagggtgca ttcattacct gcagtgcgtg gatccctgtg gtgccatcat tatcatttga
      961 agctccatct ctgaagcctc tagctttgta aggacctaca gaccacttcc caagtcattg
     1021 caggtttatt tttgtcgcat ttgagcataa cctcagattt ggcatacaat acgaatgtca
     1081 ccttgaacag cttctccttg aatgggggcc tactctcccc aggtgcagtg agcaccgtcg
     1141 tctttacatg agtgggccta caccaccctg ctgagtaaat ccatctgttc tgtttcagta
     1201 actgtggcct ctacaaatgg gaacgggacg gaatgctaaa ggtatgccat ttgcgtacta
     1261 atttttgact ccttttagtg acccatgcta ataatgtgga accatctcct attaaaatat
     1321 tttcattttt ctaggaaaag actggtccaa aaataggagg agaaactctg ttaccaagag
     1381 gagaagaaca tgctaaattt ctgaacagcc ttaaaggtgg atctggaggt ggaggttctg
     1441 gaggagaaga acttttgagc aagaattatc atcttgagaa cgaagtggct cgtcttaaga
     1501 aaggttctgg cagtggagaa gaactgcttt caaagaatta ccacctggaa aatgaggtag
     1561 ctagactgaa aaaggggagc ggaagtgggg aggagttgct gagcaaaaat tatcatttgg
     1621 agaacgaagt agcacgacta aagaaagggt ccggatcggg tgaggagtta ctctcgaaaa
     1681 attatcatct cgaaaacgaa gtggctcggc taaaaaaggg cagtggttct ggagaagagc
     1741 tattatctaa aaactaccac ctcgaaaatg aggtggcacg cttaaaaaag ggaagtggca
     1801 gtggtgaaga gctactatcc aagaattatc atcttgagaa cgaggtagcg cgtttgaaga
     1861 agggttccgg ctcaggagag gaactgctct cgaagaacta tcatcttgaa aatgaggtcg
     1921 ctcgattaaa aaagggatcg ggcagtggtg aggaactact ttcaaagaat taccacctcg
     1981 aaaacgaagt agctcgatta aagaaaggtt cagggtcggg tgaagaatta ctgagtaaaa
     2041 attatcatct ggaaaatgag gtagcgagac taaaaaaggg gagtggttct ggcgaagagt
     2101 tgctatcgaa aaattatcat cttgagaacg aagttgctag gctcaaaaag ggctcaggct
     2161 caggctaacc cgaacttcag acattttccc acagacttcc tggcctcctg tgactctgga
     2221 aagcaaagga ttggctgtgt attgtccatt gattcctgat tgacgccgtc aaaaacaaat
     2281 gcttgttaag cccataagct ttgcctgctt actttctgcc attgggttgg tttgatacca
     2341 catttaacat tgacatttaa gtggaaaacc aagttatcat tgtcttttct aagctcagtg
     2401 tggatgattg cattacttca ttcactgaag tttttgccca aaaattggaa ggtaaacaga
     2461 gagctatgtt tctgtatctt ttggttatag agtgttcact tctttatcat aacaaaattc
     2521 tagtgtttat acgaacaccc agaggcaaaa gaatttggct taattctcac tccaggtaag
     2581 tagcttaact tctgggcttc agttttctca tctgtaaaat caggaagatt ggactaagtg
     2641 atcctgaaat gtatttttta gcactggatt tctacaaata ataaaacttt cccatctaga
     2701 taatgatgat cacatagtct tgatgtacgg acattaaaag ccagatttct tcattcaatt
     2761 ctgttatctc tgttttactc tttgaaattg atcaagccac tgaatcactt tgcatttcag
     2821 tttatatata tagagagaaa gaaggtgtct gctcttacat tattgtggag ccctgtgata
     2881 gaaatatgta aaatctcata ttattttttt tttaattttt ttatttttta tgacagggtc
     2941 tcactatgtc accctggctg gagtgcagta gtgcgatcgc ggcacactgc agccttggct
     3001 tccctgggct caagcagtcc tcccacctca gtctcccaaa tagctaggac tacaggcgtg
     3061 cgtgaccaag cccagctaat ttttgcattt tttgtagaga tggggttttg ccatgttgct
     3121 caggctggtc tcaaactcct gagcactagc aatccaccca cctctatccc aatggcgcgc
     3181 cgagcttggc tcgagcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3241 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     3301 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3361 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3421 gttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3481 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3541 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3601 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3661 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3721 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3781 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3841 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3901 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3961 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4021 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4081 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4141 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4201 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4261 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4321 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttagaaaaac tcatcgagca
     4381 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     4441 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     4501 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     4561 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     4621 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     4681 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg aaacgaaata
     4741 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     4801 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     4861 ctgttttccc agggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     4921 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     4981 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     5041 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     5101 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc
     5161 gttgaatatg gctcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5221 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5281 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     5341 cctataaaaa taggcgtatc acgaggccct tttgtc
//
