LOCUS       Exported                6218 bp ds-DNA     circular SYN 22-JAN-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6218)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Feb 5, 2019 from SnapGene 4.2.6
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6218
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        129..332
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             415..420
     CDS             421..597
                     /codon_start=1
                     /label=AcrIIC3
                     /translation="MAFKRAIIFTSFNGFEKVSRTEKRRLAKIINARVSIIDEYLRAKD
                     TNASLDGQYRAFLF"
     primer_bind     complement(571..597)
                     /label=#1426
     primer_bind     complement(571..594)
                     /label=1498_MH_AcrIIC3_L58_re
     primer_bind     585..625
                     /label=#1540_MH_LOV2_F59_G_fw
     primer_bind     585..625
                     /label=#1539_MH_LOV2_F59_GS_fw
     primer_bind     585..625
                     /label=#1510_MH_LOV2_D61_fw
     primer_bind     585..625
                     /label=#1509_MH_LOV2_N60_fw
     primer_bind     585..625
                     /label=#1507_MH_LOV2_P64_fw
     primer_bind     585..625
                     /label=#1505_MH_LOV2_S63_fw
     primer_bind     585..625
                     /label=#1503_MH_LOV2_E62_fw
     primer_bind     585..625
                     /label=#1501_MH_LOV2_F59_fw
     primer_bind     597..625
                     /label=#1538_MH_LOV2_F59_GSG_fw
     CDS             601..1029
                     /codon_start=1
                     /label=AsLOV2 hs
                     /translation="LATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNLFHLQPMRDQKGDVQ
                     YFIGVQLDGTEHVRDAAEREGVMLIKKTAENIDEAAKEL"
     variation       601..1029
                     /label=Loop4
     primer_bind     601..625
                     /label=#1499_MH_LOV2_L58_fw
     primer_bind     complement(1008..1051)
                     /label=#1512_MH_LOV2_D61_re
     primer_bind     complement(1008..1051)
                     /label=#1511_MH_LOV2_E62_re
     primer_bind     complement(1008..1051)
                     /label=#1504_MH_LOV2_S63_re
     primer_bind     complement(1008..1051)
                     /label=#1502_MH_LOV2_N60_re
     primer_bind     complement(1008..1051)
                     /label=#1500_MH_LOV2_F59_re
     primer_bind     complement(1008..1030)
                     /label=#1508_MH_LOV2_A65_re
     primer_bind     complement(1008..1029)
                     /label=#1506_MH_LOV2_P64_re
     primer_bind     complement(1011..1051)
                     /label=#1543_MH_LOV2_N60_G_re
     primer_bind     complement(1011..1051)
                     /label=#1542_MH_LOV2_N60_SG_re
     primer_bind     complement(1011..1051)
                     /label=#1541_MH_LOV2_N60_GSG_re
     primer_bind     1039..1059
                     /label=#1425
     CDS             1048..1209
                     /codon_start=1
                     /label=AcrIIC3
                     /translation="AMTEFLAKLKAFAESCTGISIDAWEIEESEYVRLPVERRDFLAAA
                     NGKEIFKI"
     primer_bind     1048..1072
                     /label=1497_MH_AcrIIC3_A65_fw
     misc_feature    1249..1254
                     /label=BsmBI
     misc_feature    1294..1299
                     /label=BsmBI
     polyA_signal    1332..1521
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    1522..1556
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1602..2030
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2044..2373
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2224..2359
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2440..3234
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3408..3529
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3578..3594)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3602..3618
                     /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(3626..3656)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3671..3692
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     3792..3812
                     /label=#1506_MH_LOV2_P64_re
     primer_bind     3802..3812
                     /label=#1512_MH_LOV2_D61_re
     primer_bind     3802..3812
                     /label=#1511_MH_LOV2_E62_re
     primer_bind     3802..3812
                     /label=#1504_MH_LOV2_S63_re
     primer_bind     3802..3812
                     /label=#1502_MH_LOV2_N60_re
     primer_bind     3802..3812
                     /label=#1500_MH_LOV2_F59_re
     rep_origin      complement(3980..4565)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4736..5596)
                     /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(5597..5701)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(5967..6218,1..128)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
       61 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      121 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      181 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      241 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      301 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt
      361 actggcttat cgaaattaat acgactcact atagggagac ccaagctggc tagcgccacc
      421 atggctttca agagagctat tatatttacg tcctttaatg gttttgagaa ggtctctcga
      481 acagagaagc gacgccttgc caagataatt aacgcgaggg tctcaattat tgatgagtat
      541 ctcagagcca aggatacaaa cgcgagcctg gatggccaat accgagcctt tctgtttgga
      601 cttgctacaa ctttggagcg gattgagaag aatttcgtca taactgatcc caggttgcct
      661 gataatccta taatattcgc ctccgatagt tttcttcaac ttacggagta ctccagagag
      721 gagatactgg gaaggaattg cagattcctc caaggtccag aaactgatag agctactgtg
      781 aggaagatac gggatgccat tgacaatcaa actgaagtga cagtgcaact tatcaattac
      841 accaagtcag ggaaaaaatt ctggaacctg ttccatttgc aacccatgag ggaccaaaag
      901 ggcgatgtgc aatatttcat aggcgttcaa cttgatggta ctgaacacgt cagagatgcg
      961 gcggagcgag aaggagtgat gctgattaag aaaacagcag agaatatcga cgaagcggcg
     1021 aaagagttgg gaaacgacga aagccccgcg atgacagagt tcctcgcaaa actgaaagca
     1081 ttcgcggaga gctgcacggg aatatcaata gatgcgtggg agattgaaga gagcgagtat
     1141 gttcgactgc ccgttgaacg ccgggatttc ctggccgctg cgaatggcaa ggaaatcttc
     1201 aagatataac tcgagcggcc gccactgtgc tggatatctg cagaattaga gacgccgttg
     1261 atgttgaagt ttatataccg ttgatgttga agtcgtctct agcttaagtt taaaccgctg
     1321 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     1381 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     1441 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     1501 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     1561 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     1621 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     1681 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     1741 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     1801 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     1861 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     1921 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     1981 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     2041 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2101 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2161 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     2221 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     2281 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     2341 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     2401 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     2461 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     2521 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     2581 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     2641 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     2701 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     2761 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     2821 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     2881 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     2941 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     3001 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     3061 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     3121 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     3181 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     3241 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     3301 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     3361 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     3421 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3481 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     3541 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     3601 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     3661 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     3721 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3781 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3841 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3901 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3961 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4021 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4081 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4141 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4201 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4261 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4321 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4381 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4441 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4501 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4561 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4621 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4681 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4741 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4801 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4861 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4921 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4981 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5041 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5101 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5161 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5221 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5281 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5341 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5401 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5461 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5521 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5581 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5641 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5701 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     5761 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     5821 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     5881 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     5941 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
     6001 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     6061 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     6121 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     6181 gtaaactgcc cacttggcag tacatcaagt gtatcata
//
