LOCUS Exported 6299 bp ds-DNA circular SYN 29-AUG-2016
DEFINITION synthetic circular DNA
KEYWORDS Untitled 11
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6299)
TITLE Direct Submission
JOURNAL Exported Monday, August 29, 2016 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6299
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 141..157
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
enhancer 318..697
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 698..901
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 931..996
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/note="3xFLAG"
/translation="DYKDHDGDYKDHDIDYKDDDDK"
polyA_signal 1058..1680
/note="hGH poly(A) signal"
/note="human growth hormone polyadenylation signal"
promoter 1722..2019
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 2073..2867
/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"
polyA_signal 3519..3653
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter complement(3659..3677)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3691..3707)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3715..3731
/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)."
promoter complement(3739..3769)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 3784..3805
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4093..4681)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4852..5712)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5713..5817)
/gene="bla"
/note="AmpR promoter"
rep_origin 5844..6299
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
961 gatcatgaca tcgattacaa ggatgacgat gacaagcttg cggccgcgaa ttcatcgata
1021 gatctgatat cggtaccagt cgactctaga ggatcccggg tggcatccct gtgacccctc
1081 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
1141 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
1201 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
1261 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
1321 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
1381 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
1441 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
1501 acaggcgtga accactgctc ccttccctgt ccttctgatt ttaaaataac tataccagca
1561 ggaggacgtc cagacacagc ataggctacc tggccatgcc caaccggtgg gacatttgag
1621 ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact cagtagatgc ctgttgaatt
1681 gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
1741 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
1801 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
1861 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
1921 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
1981 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggaattgatc agcttgggat
2041 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
2101 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
2161 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
2221 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
2281 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
2341 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
2401 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
2461 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
2521 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
2581 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
2641 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
2701 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
2761 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
2821 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
2881 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
2941 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
3001 ccagcgcggg gatctcatgc tggagttctt cgcccacccc gggctcgatc ccctcgcgag
3061 ttggttcagc tgctgcctga ggctggacga cctcgcggag ttctaccggc agtgcaaatc
3121 cgtcggcatc caggaaacca gcagcggcta tccgcgcatc catgcccccg aactgcagga
3181 gtggggaggc acgatggccg ctttggtcga cccggacggg acgctcctgc gcctgataca
3241 gaacgaattg cttgcaggca tctcatgagt gtgtcttccc gttttccgcc tgaggtcact
3301 gcgtggatgg agcgctggcg cctgctgcgc gacggcgagc tgctcaccac ccactcgcca
3361 agctggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg ccgatcataa
3421 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
3481 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
3541 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
3601 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaattccc
3661 tatagtgagt cgtattaaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta
3721 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
3781 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
3841 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
3901 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
3961 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
4021 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
4081 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
4141 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
4201 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
4261 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
4321 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
4381 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
4441 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
4501 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
4561 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
4621 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
4681 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
4741 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
4801 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
4861 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
4921 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
4981 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
5041 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
5101 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
5161 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
5221 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
5281 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
5341 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
5401 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
5461 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
5521 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
5581 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
5641 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
5701 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
5761 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
5821 atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
5881 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
5941 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
6001 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
6061 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
6121 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
6181 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
6241 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattt
//