LOCUS pDonor-GDH 4016 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Encodes a mini-transposon derived from V. cholerae Tn6677 and T7-GDH
cargo gene..
ACCESSION .
VERSION .
KEYWORDS pDonor-GDH
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4016)
AUTHORS Zhang Y, Sun X, Wang Q, Xu J, Dong F, Yang S, Yang J, Zhang Z, Qian
Y, Chen J, Zhang J, Liu Y, Tao R, Jiang Y, Yang J, Yang S
TITLE Multicopy chromosomal integration using CRISPR-associated
transposases.
JOURNAL ACS Synth Biol. 2020 Jun 17. doi: 10.1021/acssynbio.0c00073.
PUBMED 32551502
REFERENCE 2 (bases 1 to 4016)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..4016
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 116..135
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 369..386
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 503..524
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 539..569
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 577..593
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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 582..604
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 601..617
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 601..617
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 806..825
/label=T7
/note="T7 promoter, forward primer"
promoter 806..824
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 825..849
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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)."
RBS 864..886
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 1689..1706
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(1759..1777)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 1773..1820
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind complement(2017..2034)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(2017..2033)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2026..2048)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(2242..2261)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2361..2383
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(2421..2439)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 2507..2611
/gene="bla"
/label=AmpR promoter
CDS 2612..3472
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(2830..2849)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3643..215
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
61 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
121 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
181 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
241 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
301 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
361 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
421 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
481 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
541 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
601 caggaaacag ctatgaccat gattacgcca agctttgttg atacaaccat aaaatgataa
661 ttacacccat aaattgataa ttatcacacc cataaattga tattgcctct tcatggtcta
721 aacttcagta agtttacgac attttcatga agaggtcatt tccggggatc cggcgtagag
781 gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat
841 aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata catatgtatc
901 cggatttaaa aggaaaagtc gtcgctatta caggagctgc ttcagggctc ggaaaggcga
961 tggccattcg cttcggcaag gagcaggcaa aagtggttat caactattat agtaataaac
1021 aagatccgaa cgaggtaaaa gaagaggtca tcaaggcggg cggtgaagct gttgtcgtcc
1081 aaggagatgt cacgaaagag gaagatgtaa aaaatatcgt gcaaacggca attaaggagt
1141 tcggcacact cgatattatg attaataatg ccggtcttga aaatcctgtg ccatctcacg
1201 aaatgccgct caaggattgg gataaagtca tcggcacgaa cttaacgggt gcctttttag
1261 gaagccgtga agcgattaaa tatttcgtag aaaacgatat caagggaaat gtcattaaca
1321 tgtccagtgt gcacgaagtg attccttggc cattatttgt ccactatgcg gcaagtaaag
1381 gcgggatgaa gctgatgaca cgcacattag cgttggaata cgcgccgaag ggcattcgcg
1441 tcaataatat tgggccaggt gcgatcaaca cgacgatcaa taaggagaaa tttgctgacc
1501 ctgaacagag agctgatgta gaaagcatga ttccaatggg atatatcggc gaaccggagg
1561 agatcgccgc agtagcagcc tggcttgctt cgaaggaagc cagctacgtc acaggcatca
1621 cgttattcgc ggacggcggt atgacacttt atccttcatt ccaggcaggc cgcggttaat
1681 gactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag
1741 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
1801 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatctaga gtccttactg
1861 cagtagtttt gctgaaatac tcgattcaca aaaatatcaa cttatggttg ttttgtgaga
1921 tatcaatata tggttgtttt gtggttaagt tgctgattat aaataattat taaatatcac
1981 tttatggttg catcaacagg taccgagctc gaattcactg gccgtcgttt tacaacgtcg
2041 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
2101 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
2161 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
2221 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
2281 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
2341 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
2401 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
2461 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
2521 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
2581 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
2641 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
2701 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
2761 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
2821 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
2881 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
2941 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
3001 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
3061 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
3121 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
3181 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
3241 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
3301 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
3361 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
3421 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
3481 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
3541 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
3601 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
3661 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
3721 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
3781 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
3841 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
3901 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
3961 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactga
//
