LOCUS       Exported                6799 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pB18cmBFP1A5
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6799)
  AUTHORS   Mena MA, Treynor TP, Mayo SL, Daugherty PS
  TITLE     Blue fluorescent proteins with enhanced brightness and 
            photostability from a structurally targeted library.
  JOURNAL   Nat Biotechnol. 2006 Dec . 24(12):1569-71.
  PUBMED    17115054
REFERENCE   2  (bases 1 to 6799)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6799
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(661..1320)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1235..1254
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1751..2206
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1838..1857)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2317..2905
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2806..2825
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3059..3076
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3091..3231
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3157..3176)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3317..3339
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3478..3497
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3677..4555)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4582..4866
                     /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)"
     primer_bind     4789..4808
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             4924..5637
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Mena et al., 2006)"
                     /label=Azurite BFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNANSHNIYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FRTAAGITHGMDELYK"
     CDS             5638..5655
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5749..5766)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5919..6005
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6097..6124
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        6143..6234
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6419..6797
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     complement(6453..6472)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gtgctgctga agttgcccgc aacagagagt ggaaccaacc ggtgatacca cgatactatg
       61 actgagagtc aacgccatga gcggcctcat ttcttattct gagttacaac agtccgcacc
      121 gctgtccggt agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
      181 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgcccaaca gtcccccggc
      241 cacggggcct gccaccatac ccacgccgaa acaagcgccc tgcaccatta tgttccggat
      301 ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg
      361 ctaaccgttt ttatcaggct ctgggaggca gaataaatga tcatatcgtc aattattacc
      421 tccacgggga gagcctgagc aaactggcct caggcatttg agaagcacac ggtcacactg
      481 cttccggtag tcaataaacc ggtaaaccag caatagacat aagcggctat ttaacgaccc
      541 tgccctgaac cgacgaccgg gtcgaatttg ctttcgaatt tctgccattc atccgcttat
      601 tatcacttat tcaggcgtag caccaggcgt ttaagggcac caataactgc cttaaaaaaa
      661 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac
      721 atggaagcca tcacagacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
      781 gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc
      841 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt
      901 ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga
      961 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt
     1021 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc
     1081 accgtctttc attgccatac ggaattccgg atgagcattc atcaggcggg caagaatgtg
     1141 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat
     1201 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg
     1261 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat
     1321 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     1381 tatttcatta tggtgaaagt tggaacctct tacgcaaact attaactggc gaactactta
     1441 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     1501 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     1561 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     1621 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     1681 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1741 agattgattt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1801 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     1861 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     1921 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     1981 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2041 tttaatagtg gactcttgtt ccaaacttga acaacactca accctatctc gggctattct
     2101 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2161 caaaaattta acgcgaattt taacaaaata ttaacgttta caatttaaaa ggatctaggt
     2221 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2281 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2341 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2401 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2461 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2521 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2581 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2641 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2701 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2761 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2821 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2881 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     2941 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3001 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3061 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     3121 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     3181 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     3241 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3301 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3361 cgcgcgaggc agcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat
     3421 acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt
     3481 gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat
     3541 gcgtccggcg tagaggatct gctcatgttt gacagcttat catcgatgca taatgtgcct
     3601 gtcaaatgga cgaagcaggg attctgcaaa ccctatgcta ctccgtcaag ccgtcaattg
     3661 tctgattcgt taccaattat gacaacttga cggctacatc attcactttt tcttcacaac
     3721 cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga
     3781 gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca
     3841 aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta
     3901 actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc
     3961 tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta
     4021 cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca
     4081 attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt
     4141 taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga
     4201 accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac
     4261 gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa
     4321 tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat
     4381 ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg
     4441 gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca
     4501 gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt
     4561 tcatactccc gccattcaga gaagaaacca attgtccata ttgcatcaga cattgccgtc
     4621 actgcgtctt ttactggctc ttctcgctaa ccaaaccggt aaccccgctt attaaaagca
     4681 ttctgtaaca aagcgggacc aaagccatga caaaaacgcg taacaaaagt gtctataatc
     4741 acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat gccatagcat
     4801 ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc tctactgttt
     4861 ctccataccc gtttttttgg gctagcgaat tcgagctcgg ccacgaaggc caggagtgaa
     4921 acgatgtcta aaggtgaaga attatttact ggtgttgtcc caattttggt tgaattagat
     4981 ggtgatgtta atggtcacaa attttctgtc tccggtgaag gtgaaggtga tgctacgtac
     5041 ggtaaattga ccttaaaatt tatttgtact actggtaaat tgccagttcc atggccaacc
     5101 ttagtaacta ctttgagcca tggtgttcaa tgtttttcta gatacccaga tcatatgaaa
     5161 caacatgact ttttcaagtc tgccatgcca gaaggttatg ttcaagaaag aactattttt
     5221 ttcaaagatg acggtaacta caagaccaga gctgaagtca agtttgaagg tgatacctta
     5281 gttaatagaa tcgaattaaa aggtattgat tttaaagaag atggtaacat tttaggtcac
     5341 aaattggaat acaacgccaa ctctcacaat atatacatca tggctgacaa acaaaagaat
     5401 ggtatcaaag cgaacttcaa aattagacac aacattgaag atggttctgt tcaattagct
     5461 gaccattatc aacaaaatac tccaattggt gatggtccag tcttgttacc agacaaccat
     5521 tacttatcca cccaatcagc cttatccaaa gatccaaacg aaaagagaga ccacatggtc
     5581 ctgttagaat ttaggactgc tgctggtatt acccatggta tggatgaatt gtacaaacac
     5641 caccaccacc accactaagg ccaaggtggc cggtacccgg ggatcctcta gagtcgacct
     5701 gcaggcatgc aagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat
     5761 taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt
     5821 ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt
     5881 ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
     5941 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     6001 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
     6061 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
     6121 tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg
     6181 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6241 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     6301 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     6361 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6421 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     6481 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6541 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6601 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6661 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6721 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     6781 gcaatggcaa caacgttgc
//
