LOCUS Exported 6598 bp ds-DNA circular SYN 26-SEP-2017
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCMV6-AC-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6598)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 26, 2017 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6598
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 166..182
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
enhancer 343..722
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 723..926
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 877..897
/label=CMV-F
promoter 952..970
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 979..1087
/label=MCS
CDS 1088..1783
/codon_start=1
/product="improved green fluorescent protein from
Pontellina plumata (Evdokimov et al., 2006)"
/label=TurboGFP
/note="mammalian codon-optimized"
/translation="MESDESGLPAMEIECRITGTLNGVEFELVGGGEGTPEQGRMTNKM
KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
HVSFSYRYEAGRVIGDFKVMGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNDLDGSFTR
TFSLRDGGYYSSVVDSHMHFKSAIHPSILQNGGPMFAFRRVEEDHSNTELGIVEYQHAF
KTPDADAGEE"
primer_bind complement(1172..1191)
/label=TurboGFP-R
primer_bind complement(1806..1822)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
polyA_signal 1833..2455
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
promoter 2484..2813
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2664..2799
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2880..3674
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=Neo
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 3848..3969
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4018..4034)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 4042..4058
/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)."
promoter complement(4066..4096)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4111..4132
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4420..5005)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5176..6036)
/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"
promoter complement(6037..6141)
/gene="bla"
/label=AmpR promoter
rep_origin 6168..25
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg
1021 cgatcgccgg cgcgccagat ctcaagctta actagttagc ggaccgacgc gtacgcggcc
1081 gctcgagatg gagagcgacg agagcggcct gcccgccatg gagatcgagt gccgcatcac
1141 cggcaccctg aacggcgtgg agttcgagct ggtgggcggc ggagagggca cccccgagca
1201 gggccgcatg accaacaaga tgaagagcac caaaggcgcc ctgaccttca gcccctacct
1261 gctgagccac gtgatgggct acggcttcta ccacttcggc acctacccca gcggctacga
1321 gaaccccttc ctgcacgcca tcaacaacgg cggctacacc aacacccgca tcgagaagta
1381 cgaggacggc ggcgtgctgc acgtgagctt cagctaccgc tacgaggccg gccgcgtgat
1441 cggcgacttc aaggtgatgg gcaccggctt ccccgaggac agcgtgatct tcaccgacaa
1501 gatcatccgc agcaacgcca ccgtggagca cctgcacccc atgggcgata acgatctgga
1561 tggcagcttc acccgcacct tcagcctgcg cgacggcggc tactacagct ccgtggtgga
1621 cagccacatg cacttcaaga gcgccatcca ccccagcatc ctgcagaacg ggggccccat
1681 gttcgccttc cgccgcgtgg aggaggatca cagcaacacc gagctgggca tcgtggagta
1741 ccagcacgcc ttcaagaccc cggatgcaga tgccggtgaa gaaagagttt aaacggccgg
1801 ccgcggtcat agctgtttcc tgaacagatc ccgggtggca tccctgtgac ccctccccag
1861 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat
1921 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg
1981 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg
2041 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc
2101 aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc
2161 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct
2221 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg
2281 cgtgaaccac tgctcccttc cctgtccttc tgattttaaa ataactatac cagcaggagg
2341 acgtccagac acagcatagg ctacctggcc atgcccaacc ggtgggacat ttgagttgct
2401 tgcttggcac tgtcctctca tgcgttgggt ccactcagta gatgcctgtt gaattgggta
2461 cgcggccagc ttggctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
2521 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
2581 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
2641 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
2701 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
2761 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
2821 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
2881 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
2941 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
3001 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
3061 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
3121 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
3181 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
3241 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
3301 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
3361 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
3421 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
3481 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggccgaccgc tatcaggaca
3541 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
3601 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
3661 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
3721 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
3781 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
3841 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
3901 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
3961 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
4021 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
4081 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
4141 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
4201 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
4261 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
4321 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
4381 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
4441 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
4501 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
4561 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
4621 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
4681 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
4741 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
4801 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
4861 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
4921 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
4981 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
5041 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
5101 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
5161 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
5221 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
5281 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
5341 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
5401 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
5461 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
5521 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
5581 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
5641 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
5701 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
5761 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
5821 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
5881 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
5941 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
6001 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
6061 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
6121 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag
6181 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
6241 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
6301 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
6361 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
6421 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
6481 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
6541 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaatttt
//