LOCUS Exported 5144 bp ds-DNA circular SYN 10-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pBBR1MCS-2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5144)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Saturday, September 10, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..5144
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 458..1252
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
protein_bind 1657..1678
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1693..1723
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 1731..1747
/bound_moiety="lac repressor encoded by lacI"
/note="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)."
primer_bind 1755..1771
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS 1767..2132
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/note="lacZ-alpha"
/translation="MTMITPSAQLTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
LERPPPRWSSNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
TDRPSQQLRSLNGEWKL"
promoter 1792..1810
/note="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
misc_feature 1823..1930
/note="MCS"
/note="pBluescript multiple cloning site"
primer_bind 1840..1856
/note="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1890..1906)
/note="SK primer"
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(1939..1957)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1967..1983)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(2690..3352)
/codon_start=1
/product="replication protein for the broad-host-range
plasmid pBBR1 from Bordetella bronchiseptica"
/note="pBBR1 Rep"
/translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
rep_origin 3353..4122
/note="pBBR1 oriV"
/note="replication origin of the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica; requires the pBBR1
Rep protein for replication"
ORIGIN
1 accttcggga gcgcctgaag cccgttctgg acgccctggg gccgttgaat cgggatatgc
61 aggccaaggc cgccgcgatc atcaaggccg tgggcgaaaa gctgctgacg gaacagcggg
121 aagtccagcg ccagaaacag gcccagcgcc agcaggaacg cgggcgcgca catttccccg
181 aaaagtgcca cctgggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag
241 cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt
301 tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa
361 gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc
421 aagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
481 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
541 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
601 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
661 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
721 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
781 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
841 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
901 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
961 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
1021 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
1081 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
1141 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
1201 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
1261 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
1321 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
1381 atcctccagc gcggggatct catgctggag ttcttcgccc acccccatgg gcaaatatta
1441 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
1501 tggcttccat gtcggcagaa tgcttaatga attacaacag tttttatgca tgcgcccaat
1561 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
1621 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
1681 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
1741 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
1801 tcactaaagg gaacaaaagc tgggtaccgg gccccccctc gaggtcgacg gtatcgataa
1861 gcttgatatc gaattcctgc agcccggggg atccactagt tctagagcgg ccgccaccgc
1921 ggtggagctc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
1981 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
2041 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
2101 tgcgcagcct gaatggcgaa tggaaattgt aagcgttaat attttgttaa aattcgcgtt
2161 aaatttttgt taaatcagct cattttttaa ccaataggcc gactgcgatg agtggcaggg
2221 cggggcgtaa tttttttaag gcagttattg gtgcccttaa acgcctggtg ctacgcctga
2281 ataagtgata ataagcggat gaatggcaga aattcgaaag caaattcgac ccggtcgtcg
2341 gttcagggca gggtcgttaa atagccgctt atgtctattg ctggtttacc ggtttattga
2401 ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg aggccagttt gctcaggctc
2461 tccccgtgga ggtaataatt gacgatatga tcatttattc tgcctcccag agcctgataa
2521 aaacggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct
2581 ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcga ggcggctaca
2641 gccgatagtc tggaacagcg cacttacggg ttgctgcgca acccaagtgc taccggcgcg
2701 gcagcgtgac ccgtgtcggc ggctccaacg gctcgccatc gtccagaaaa cacggctcat
2761 cgggcatcgg caggcgctgc tgcccgcgcc gttcccattc ctccgtttcg gtcaaggctg
2821 gcaggtctgg ttccatgccc ggaatgccgg gctggctggg cggctcctcg ccggggccgg
2881 tcggtagttg ctgctcgccc ggatacaggg tcgggatgcg gcgcaggtcg ccatgcccca
2941 acagcgattc gtcctggtcg tcgtgatcaa ccaccacggc ggcactgaac accgacaggc
3001 gcaactggtc gcggggctgg ccccacgcca cgcggtcatt gaccacgtag gccgacacgg
3061 tgccggggcc gttgagcttc acgacggaga tccagcgctc ggccaccaag tccttgactg
3121 cgtattggac cgtccgcaaa gaacgtccga tgagcttgga aagtgtcttc tggctgacca
3181 ccacggcgtt ctggtggccc atctgcgcca cgaggtgatg cagcagcatt gccgccgtgg
3241 gtttcctcgc aataagcccg gcccacgcct catgcgcttt gcgttccgtt tgcacccagt
3301 gaccgggctt gttcttggct tgaatgccga tttctctgga ctgcgtggcc atgcttatct
3361 ccatgcggta gggtgccgca cggttgcggc accatgcgca atcagctgca acttttcggc
3421 agcgcgacaa caattatgcg ttgcgtaaaa gtggcagtca attacagatt ttctttaacc
3481 tacgcaatga gctattgcgg ggggtgccgc aatgagctgt tgcgtacccc ccttttttaa
3541 gttgttgatt tttaagtctt tcgcatttcg ccctatatct agttctttgg tgcccaaaga
3601 agggcacccc tgcggggttc ccccacgcct tcggcgcggc tccccctccg gcaaaaagtg
3661 gcccctccgg ggcttgttga tcgactgcgc ggccttcggc cttgcccaag gtggcgctgc
3721 ccccttggaa cccccgcact cgccgccgtg aggctcgggg ggcaggcggg cgggcttcgc
3781 cttcgactgc ccccactcgc ataggcttgg gtcgttccag gcgcgtcaag gccaagccgc
3841 tgcgcggtcg ctgcgcgagc cttgacccgc cttccacttg gtgtccaacc ggcaagcgaa
3901 gcgcgcaggc cgcaggccgg aggcttttcc ccagagaaaa ttaaaaaaat tgatggggca
3961 aggccgcagg ccgcgcagtt ggagccggtg ggtatgtggt cgaaggctgg gtagccggtg
4021 ggcaatccct gtggtcaagc tcgtgggcag gcgcagcctg tccatcagct tgtccagcag
4081 ggttgtccac gggccgagcg aagcgagcca gccggtggcc gctcgcggcc atcgtccaca
4141 tatccacggg ctggcaaggg agcgcagcga ccgcgcaggg cgaagcccgg agagcaagcc
4201 cgtagggcgc cgcagccgcc gtaggcggtc acgactttgc gaagcaaagt ctagtgagta
4261 tactcaagca ttgagtggcc cgccggaggc accgccttgc gctgcccccg tcgagccggt
4321 tggacaccaa aagggagggg caggcatggc ggcatacgcg atcatgcgat gcaagaagct
4381 ggcgaaaatg ggcaacgtgg cggccagtct caagcacgcc taccgcgagc gcgagacgcc
4441 caacgctgac gccagcagga cgccagagaa cgagcactgg gcggccagca gcaccgatga
4501 agcgatgggc cgactgcgcg agttgctgcc agagaagcgg cgcaaggacg ctgtgttggc
4561 ggtcgagtac gtcatgacgg ccagcccgga atggtggaag tcggccagcc aagaacagca
4621 ggcggcgttc ttcgagaagg cgcacaagtg gctggcggac aagtacgggg cggatcgcat
4681 cgtgacggcc agcatccacc gtgacgaaac cagcccgcac atgaccgcgt tcgtggtgcc
4741 gctgacgcag gacggcaggc tgtcggccaa ggagttcatc ggcaacaaag cgcagatgac
4801 ccgcgaccag accacgtttg cggccgctgt ggccgatcta gggctgcaac ggggcatcga
4861 gggcagcaag gcacgtcaca cgcgcattca ggcgttctac gaggccctgg agcggccacc
4921 agtgggccac gtcaccatca gcccgcaagc ggtcgagcca cgcgcctatg caccgcaggg
4981 attggccgaa aagctgggaa tctcaaagcg cgttgagacg ccggaagccg tggccgaccg
5041 gctgacaaaa gcggttcggc aggggtatga gcctgcccta caggccgccg caggagcgcg
5101 tgagatgcgc aagaaggccg atcaagccca agagacggcc cgag
//