LOCUS pBbB8k-GFP 4480 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pBbB8k-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4480)
AUTHORS Lee TS, Krupa RA, Zhang F, Hajimorad M, Holtz WJ, Prasad N, Lee SK,
Keasling JD
TITLE BglBrick vectors and datasheets: A synthetic biology platform for
gene expression.
JOURNAL J Biol Eng. 2011 Sep 20;5:12.
PUBMED 21933410
REFERENCE 2 (bases 1 to 4480)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4480
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(7..885)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
promoter 912..1196
/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 1119..1138
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
CDS 1246..1962
/codon_start=1
/product="GFP variant optimized for excitation by UV light"
/label=GFPuv
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
primer_bind complement(1276..1304)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind 1899..1919
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
terminator 1995..2066
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2082..2109
/label=T7Te terminator
/note="phage T7 early transcription terminator"
CDS complement(2176..2838)
/codon_start=1
/product="replication protein for the broad-host-range
plasmid pBBR1 from Bordetella bronchiseptica"
/label=pBBR1 Rep
/translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVLAYVV
NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
terminator 3425..3519
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(3550..4344)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3661..3680)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 4271..4290
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
ORIGIN
1 gacgtcttat gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa
61 ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc
121 aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt
181 cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg
241 gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc
301 aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc
361 catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag
421 cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg
481 cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt
541 ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac
601 ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg
661 cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac
721 cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat
781 aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt
841 aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc
901 gccattcaga gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt
961 ttactggctc ttctcgctaa ccaaaccggt aaccccgctt attaaaagca ttctgtaaca
1021 aagcgggacc aaagccatga caaaaacgcg taacaaaagt gtctataatc acggcagaaa
1081 agtccacatt gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat
1141 aagattagcg gattctacct gacgcttttt atcgcaactc tctactgttt ctccataccc
1201 gtttttttgg gaattcaaaa gatcttttaa gaaggagata tacatatgag taaaggagaa
1261 gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac
1321 aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa
1381 tttatttgca ctactggaaa actacctgtt ccgtggccaa cacttgtcac tactttctct
1441 tatggtgttc aatgcttttc ccgttatccg gatcacatga aacggcatga ctttttcaag
1501 agtgccatgc ccgaaggtta tgtacaggaa cgcactatat ctttcaaaga tgacgggaac
1561 tacaagacgc gtgctgaagt caagtttgaa ggtgataccc ttgttaatcg tatcgagtta
1621 aaaggtattg attttaaaga agatggaaac attctcggac acaaactgga gtacaactat
1681 aactcacaca atgtatacat cacggcagac aaacaaaaga atggaatcaa agctaacttc
1741 aaaattcgcc acaacattga agatggctcc gttcaactag cagaccatta tcaacaaaat
1801 actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct
1861 gccctttcga aagatcccaa cgaaaagcgt gaccacatgg tccttcttga gtttgtaact
1921 gctgctggga ttacacatgg catggatgag ctctacaaat aaggatccaa actcgagtaa
1981 ggatctccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
2041 tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc
2101 ctttctgcgt ttatacctag gctacagccg atagtctgga acagcgcact tacgggttgc
2161 tgcgcaaccc aagtgctacc ggcgcggcag cgtgacccgt gtcggcggct ccaacggctc
2221 gccatcgtcc agaaaacacg gctcatcggg catcggcagg cgctgctgcc cgcgccgttc
2281 ccattcctcc gtttcggtca aggctggcag gtctggttcc atgcccggaa tgccgggctg
2341 gctgggcggc tcctcgccgg ggccggtcgg tagttgctgc tcgcccggat acagggtcgg
2401 gatgcggcgc aggtcgccat gccccaacag cgattcgtcc tggtcgtcgt gatcaaccac
2461 cacggcggca ctgaacaccg acaggcgcaa ctggtcgcgg ggctggcccc acgccacgcg
2521 gtcattgacc acgtaggcca acacggtgcc ggggccgttg agcttcacga cggagatcca
2581 gcgctcggcc accaagtcct tgactgcgta ttggaccgtc cgcaaagaac gtccgatgag
2641 cttggaaagt gtcttctggc tgaccaccac ggcgttctgg tggcccatct gcgccacgag
2701 gtgatgcagc agcattgccg ccgtgggttt cctcgcaata agcccggccc acgcctcatg
2761 cgctttgcgt tccgtttgca cccagtgacc gggcttgttc ttggcttgaa tgccgatttc
2821 tctggactgc gtggccatgc ttatctccat gcggtagggg tgccgcacgg ttgcggcacc
2881 atgcgcaatc agctgcaact tttcggcagc gcgacaacaa ttatgcgttg cgtaaaagtg
2941 gcagtcaatt acagattttc tttaacctac gcaatgagct attgcggggg gtgccgcaat
3001 gagctgttgc gtacccccct tttttaagtt gttgattttt aagtctttcg catttcgccc
3061 tatatctagt tctttggtgc ccaaagaagg gcacccctgc ggggttcccc cacgccttcg
3121 gcgcggctcc ccctccggca aaaagtggcc cctccggggc ttgttgatcg actgcgcggc
3181 cttcggcctt gcccaaggtg gcgctgcccc cttggaaccc ccgcactcgc cgccgtgagg
3241 ctcggggggc aggcgggcgg gcttcgccct tcgactgccc ccactcgcat aggcttgggt
3301 cgttccaggc gcgtcaaggc caagccgctg cgcggtcgct gcgcgagcct tgacccgcct
3361 tccacttggt gtccaaccgg caagcgaagc gcgcaggccg caggccggag gcactagtgc
3421 ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat
3481 ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc tcgaacccca
3541 gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg
3601 agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc
3661 aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca
3721 gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc
3781 atgggtcacg acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc
3841 ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc
3901 catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc
3961 cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg
4021 agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct
4081 tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca
4141 cgatagccgc gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac
4201 aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat
4261 tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc
4321 gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcagatca
4381 tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta ctttgcaggg
4441 cttcccaacc ttaccagagg gcgccccagc tggcaattcc
//
