LOCUS Exported 4291 bp ds-DNA circular SYN 19-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4291)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Monday, September 19, 2016 from SnapGene Viewer 3.2.1
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..4291
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 268..295
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 387..473
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 537..553
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 570..801
/gene="mutant version of attP"
/bound_moiety="BP Clonase(TM)"
/note="attP1"
/note="recombination site for the Gateway(R) BP reaction"
CDS complement(1197..1502)
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/note="ccdB"
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
CDS complement(1847..2506)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(2507..2609)
/note="cat promoter"
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
protein_bind complement(2754..2985)
/gene="mutant version of attP"
/bound_moiety="BP Clonase(TM)"
/note="attP2"
/note="recombination site for the Gateway(R) BP reaction"
promoter complement(3004..3022)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3027..3043)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(3111..3485)
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/note="BleoR"
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
promoter complement(3504..3551)
/note="EM7 promoter"
/note="synthetic bacterial promoter "
rep_origin 3641..4229
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
661 agctgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa
721 aaaacagact acataatact gtaaaacaca acatatccag tcactatgaa tcaactactt
781 agatggtatt agtgacctgt agtcgaccga cagccttcca aatgttcttc gggtgatgct
841 gccaacttag tcgaccgaca gccttccaaa tgttcttctc aaacggaatc gtcgtatcca
901 gcctactcgc tattgtcctc aatgccgtat taaatcataa aaagaaataa gaaaaagagg
961 tgcgagcctc ttttttgtgt gacaaaataa aaacatctac ctattcatat acgctagtgt
1021 catagtcctg aaaatcatct gcatcaagaa caatttcaca actcttatac ttttctctta
1081 caagtcgttc ggcttcatct ggattttcag cctctatact tactaaacgt gataaagttt
1141 ctgtaatttc tactgtatcg acctgcagac tggctgtgta taagggagcc tgacatttat
1201 attccccaga acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg gctgagatca
1261 gccacttctt ccccgataac ggagaccggc acactggcca tatcggtggt catcatgcgc
1321 cagctttcat ccccgatatg caccaccggg taaagttcac gggagacttt atctgacagc
1381 agacgtgcac tggccagggg gatcaccatc cgtcgcccgg gcgtgtcaat aatatcactc
1441 tgtacatcca caaacagacg ataacggctc tctcttttat aggtgtaaac cttaaactgc
1501 atttcaccag cccctgttct cgtcagcaaa agagccgttc atttcaataa accgggcgac
1561 ctcagccatc ccttcctgat tttccgcttt ccagcgttcg gcacgcagac gacgggcttc
1621 attctgcatg gttgtgctta ccagaccgga gatattgaca tcatatatgc cttgagcaac
1681 tgatagctgt cgctgtcaac tgtcactgta atacgctgct tcatagcata cctctttttg
1741 acatacttcg ggtatacata tcagtatata ttcttatacc gcaaaaatca gcgcgcaaat
1801 acgcatactg ttatctggct tttagtaagc cggatccacg cggcgtttac gccccgccct
1861 gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac
1921 agacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
1981 atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa
2041 aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt
2101 tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa
2161 actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat
2221 ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg
2281 ccatacggaa ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat
2341 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg
2401 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc
2461 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag
2521 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
2581 gaaagttgga acctcttacg tgccgatcaa cgtctcattt tcgccaaaag ttggcccagg
2641 gcttcccggt atcaacaggg acaccaggat ttatttattc tgcgaagtga tcttccgtca
2701 caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc caacttagtc gactacaggt
2761 cactaatacc atctaagtag ttgattcata gtgactggat atgttgtgtt ttacagtatt
2821 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg
2881 tttctcgttc agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt
2941 gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata
3001 tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt
3061 ctcaaaatct ctgatgttac attgcacaag ataaaataat atcatcatga tcagtcctgc
3121 tcctcggcca cgaagtgcac gcagttgccg gccgggtcgc gcagggcgaa ctcccgcccc
3181 cacggctgct cgccgatctc ggtcatggcc ggcccggagg cgtcccggaa gttcgtggac
3241 acgacctccg accactcggc gtacagctcg tccaggccgc gcacccacac ccaggccagg
3301 gtgttgtccg gcaccacctg gtcctggacc gcgctgatga acagggtcac gtcgtcccgg
3361 accacaccgg cgaagtcgtc ctccacgaag tcccgggaga acccgagccg gtcggtccag
3421 aactcgaccg ctccggcgac gtcgcgcgcg gtgagcaccg gaacggcact ggtcaacttg
3481 gccatggttt agttcctcac cttgtcgtat tatactatgc cgatatacta tgccgatgat
3541 taattgtcaa cacgtgctga tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
3601 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
3661 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
3721 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
3781 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
3841 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
3901 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
3961 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
4021 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
4081 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
4141 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
4201 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
4261 tggccttttg ctggcctttt gctcacatgt t
//