LOCUS Exported 6817 bp ds-DNA circular SYN 30-AUG-2016
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS pBC-hygro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6817)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2016-02-20 from MiaoLingBio http://www.miaolingbio.com
REFERENCE 2 (bases 1 to 6817)
AUTHORS .
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6817)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Tuesday, August 30, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6817
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(3..458)
/direction=LEFT
/note="f1 ori"
/note="f1 ori;f1 ori;f1 bacteriophage origin of
replication; arrow indicates direction of (+) strand
synthesis"
CDS complement(241..816)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/note="lacZ"
/note="lacZ fragment;lacZ-alpha"
/translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
TDRPSQQLRSLNGEWDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG
FPRQALNRGLPLGFRFSALRHLDPKKLD"
primer_bind 600..616
/note="M13 fwd"
/note="M13 fwd;M13 fwd;common sequencing primer, one of
multiple similar variants"
promoter 626..644
/note="T7 promoter"
/note="T7 promoter;T7 promoter;promoter for bacteriophage
T7 RNA polymerase"
misc_feature 653..760
/note="MCS"
/note="MCS;MCS;pBluescript multiple cloning site"
primer_bind 670..686
/note="KS primer"
/note="KS primer;KS primer;common sequencing primer, one of
multiple similar variants"
primer_bind complement(720..736)
/note="SK primer"
/note="SK primer;SK primer;common sequencing primer, one of
multiple similar variants"
promoter complement(773..791)
/note="T3 promoter"
/note="T3 promoter;T3 promoter;promoter for bacteriophage
T3 RNA polymerase"
primer_bind complement(812..828)
/note="M13 rev"
/note="M13 rev;M13 rev;common sequencing primer, one of
multiple similar variants"
protein_bind 836..852
/bound_moiety="lac repressor encoded by lacI"
/note="laco"
/note="lac repressor encoded by lacI lac operator;lac
operator;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(860..890)
/note="lac promoter"
/note="lac promoter;lac promoter;promoter for the E. coli
lac operon"
protein_bind 905..926
/bound_moiety="E. coli catabolite activator protein"
/note="CAP"
/note="E. coli catabolite activator protein CAP bindin;CAP
binding site;CAP binding activates transcription in the
presence of cAMP."
rep_origin complement(1214..1802)
/direction=LEFT
/note="ori"
/note="ori;ori;high-copy-number ColE1/pMB1/pBR322/pUC
origin of replication"
promoter 2022..2124
/note="cat promoter"
/note="cat promoter;cat promoter;promoter of the E. coli
cat gene encoding chloramphenicol acetyltransferase"
CDS 2125..2784
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="Chl"
/note="cat;CmR;confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
misc_feature 3732..4535
/note="CPC-1 Promoter"
CDS 4541..5566
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/note="aph"
/note="aph(4)-Ia;HygR;confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
misc_feature 5567..6501
/note="trpC Terminator"
promoter complement(6689..6794)
/gene="bla"
/note="bla bla AmpR promoter"
/note="bla AmpR promoter;AmpR promoter"
ORIGIN
1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
661 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gaattcctgc agcccggggg
721 atccactagt tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt
781 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
841 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
901 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
961 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
1021 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
1081 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
1141 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
1201 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
1261 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
1321 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
1381 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
1441 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
1501 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
1561 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
1621 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
1681 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
1741 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
1801 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
1861 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttcgaccgaa
1921 taaatacctg tgacggaaga tcacttcgca gaataaataa atcctggtgt ccctgttgat
1981 accgggaagc cctgggccaa cttttggcga aaatgagacg ttgatcggca cgtaagaggt
2041 tccaactttc accataatga aataagatca ctaccgggcg tattttttga gttgtcgaga
2101 ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat
2161 atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc
2221 tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag
2281 cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa
2341 ttacgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac
2401 accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat
2461 ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc
2521 tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt
2581 ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc
2641 atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat
2701 catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc
2761 gatgagtggc agggcggggc gtaatttttt taaggcagtt attggtgccc ttaaacgcct
2821 ggttgctacg cctgaataag tgataataag cggatgaatg gcagaaattc gaaagcaaat
2881 tcgacccggt cgtcggttca gggcagggtc gttaaatagc cgcttatgtc tattgctggt
2941 ttaccggttt attgactacc ggaagcagtg tgaccgtgtg cttctcaaat gcctgaggcc
3001 agtttgctca ggctctcccc gtggaggtaa taattgacga tatgatcctt tttttctgat
3061 caaaagtgct catcattgga aaaccgagat gcgccgcgtg cggctgctgg agatggcgga
3121 cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg
3181 attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag
3241 gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg
3301 gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc
3361 gtgacgatca gcggtccaat gatcgaagtt aggctggtaa gagccgcgag cgatccttga
3421 agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc
3481 ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgag
3541 ctttgaagtt gctgcaagct ggcttcaagc catcccatcc gaatgtgatg gatgcgttct
3601 ttctgggccg ttgcgacttt ggggatcgtc tttcccgcgc ccttggttgg aggccctgtc
3661 tccggtgtcc cttgtccctt ccaggcaagc gagcgaggtc cattcagatg gtgctccatc
3721 agcgttggct ttccgtctcc attggctctt ggcaattcgg tcagcggggc tgactgcctc
3781 aggtggggca gtgctagtgt gtgtaccgac ccgcaggatt ggtgctttgc ccagagctct
3841 acagaatagc gcgcgcatcc atatgttagt tctgcaattt tcttgtatcg gtgctgtgac
3901 tcatacttcc ccctttggct ggccttgcgg caaccaataa gaacgcacag tgaaatcttg
3961 cgggtgggga gtggatccat ggcgcctgca ttggcttggg gacgcgcact gtcgcacact
4021 tccatctgac ctttcagaag ggtttcgtgg tgggcaagga ccaaccggtt gcgcggccgt
4081 gcgtgggtgc ctcgcccggc actgccaggg ccactgcagt ggcagtttgc tgcctgatac
4141 aaaatccttc cctccgccca gttttccctc tttgaccttc ctttctcttc tctgcaacca
4201 aatccaccct atcaaaccaa aacagtatct cgaccgaggt atcaacctga atcagcaaca
4261 tcgtagccag catttgtctc cgtctctgca gaaccagcga gttgcaaaca ttatccaggc
4321 aacagggcac caactcactt cttcggcttt caccaatcgg tacagctctt ctcagaactc
4381 gcgtccgcaa cagttctacg cttcctcagc accttcttca gcttcaatcc tgaacactca
4441 gaaccgcgca cagcagcgcc ctcctgttcc cttgtttccc aaaagtaccg gtagtatttc
4501 gcacggaaag cagggcaaca agatgttctc aggtacccat atgaaaaagc ctgaactcac
4561 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
4621 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
4681 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
4741 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct
4801 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
4861 actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct
4921 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
4981 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
5041 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
5101 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
5161 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
5221 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
5281 ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct
5341 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
5401 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
5461 acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga
5521 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagagta gatgccgacc
5581 gggatccact taacgttact gaaatcatca aacagcttga cgaatctgga tataagatcg
5641 ttggtgtcga tgtcagctcc ggagttgaga caaatggtgt tcaggatctc gataagatac
5701 gttcatttgt ccaagcagca aagagtgcct tctagtgatt taatagctcc atgtcaacaa
5761 gaataaaacg cgtttcgggt ttacctcttc cagatacagc tcatctgcaa tgcattaatg
5821 cattggacct cgcaacccta gtacgccctt caggctccgg cgaagcagaa gaatagctta
5881 gcagagtcta ttttcatttt cgggagacga gatcaagcag atcaacggtc gtcaagagac
5941 ctacgagact gaggaatccg ctcttggctc cacgcgacta tatatttgtc tctaattgta
6001 ctttgacatg ctcctcttct ttactctgat agcttgacta tgaaaattcc gtcaccagcc
6061 cctgggttcg caaagataat tgcactgttt cttccttgaa ctctcaagcc tacaggacac
6121 acattcatcg taggtataaa cctcgaaaat cattcctact aagatgggta tacaatagta
6181 accatggttg cctagtgaat gctccgtaac acccaatacg ccggccgaaa cttttttaca
6241 actctcctat gagtcgttta cccagaatgc acaggtacac ttgtttagag gtaatccttc
6301 tttctagcta gaggatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag
6361 gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact
6421 tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac
6481 tgttgggcgc catctccttg cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
6541 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
6601 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
6661 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
6721 atcaagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
6781 ataggggttc cgcgcacatt tccccgaaaa gtgccac
//