LOCUS       Exported                7322 bp DNA     circular SYN 03-NOV-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7322)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 12, 2022 from SnapGene 6.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7322
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             917..943
                     /codon_start=1
                     /label=PKA-Myristoylation
                     /note="color: #ff7e79"
                     /translation="MGNAAAAKK"
     CDS             944..1150
                     /codon_start=1
                     /label=DnaJB1 (1-70)
                     /note="color: #945200"
                     /translation="GKDYYQTLGLARGASDEEIKRAYRRQALRYHPDKNKEPGAEEKFK
                     EIAEAYDVLSDPRKREIFDRYGEE"
     CDS             1151..2161
                     /codon_start=1
                     /label=PKA-Calpha (16-351)
                     /note="color: #942193"
                     /translation="VKEFLAKAKEDFLKKWESPAQNTAHLDQFERIKTLGTGSFGRVML
                     VKHKETGNHYAMKILDKQKVVKLKQIEHTLNEKRILQAVNFPFLVKLEFSFKDNSNLYM
                     VMEYVPGGEMFSHLRRIGRFSEPHARFYAAQIVLTFEYLHSLDLIYRDLKPENLLIDQQ
                     GYIHVTDFGFAKRVKGRTWTLCGTPEYLAPEIILSKGYNKAVDWWALGVLIYEMAAGYP
                     PFFADQPIQILYEKIVSGKVRFPSHFSSDLKDLLRNLLQVDLTKRFGNLKNGVNDIKNH
                     KWFATTDWIAIYQRKVEAPFIPKFKGPGDTSNFDDYEEEEIRVSINEKCGKEFSEF"
     CDS             2174..2884
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /note="color: #ff0000"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     polyA_signal    2922..3146
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      3192..3620
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3634..3963
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      3814..3949
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             4030..4824
                     /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))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4998..5119
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(5168..5184)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    5192..5208
                     /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)."
                     /note="color: #31849b"
     promoter        complement(5216..5246)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 5216 .. 5222 / #ffffff / -10
                        2: 5223 .. 5240 / #ffffff
                        3: 5241 .. 5246 / #ffffff / -35"
     protein_bind    5261..5282
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(5570..6155)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6326..7186)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 6326 .. 7117 / #ccffcc
                        2: 7118 .. 7186 / #ccffcc / signal sequence
                     Cleavage site after base 7117"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7187..7291)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttatgg gcaacgccgc cgccgccaag aagggtaaag actactacca
      961 gacgttgggc ctggcccgcg gcgcgtcgga cgaggagatc aagcgggcct accgccgcca
     1021 ggcgctgcgc taccacccgg acaagaacaa ggagcccggc gccgaggaga agttcaagga
     1081 gatcgctgag gcctacgacg tgctcagcga cccgcgcaag cgcgagatct tcgaccgcta
     1141 cggggaggaa gtgaaagaat tcttagccaa agccaaagaa gattttctta aaaaatggga
     1201 aagtcccgct cagaacacag cccacttgga tcagtttgaa cgaatcaaga ccctcggcac
     1261 gggctccttc gggcgggtga tgctggtgaa acacaaggag accgggaacc actatgccat
     1321 gaagatcctc gacaaacaga aggtggtgaa actgaaacag atcgaacaca ccctgaatga
     1381 aaagcgcatc ctgcaagctg tcaactttcc gttcctcgtc aaactcgagt tctccttcaa
     1441 ggacaactca aacttataca tggtcatgga gtacgtgccc ggcggggaga tgttctcaca
     1501 cctacggcgg atcggaaggt tcagtgagcc ccatgcccgt ttctacgcgg cccagatcgt
     1561 cctgaccttt gagtatctgc actcgctgga tctcatctac agggacctga agccggagaa
     1621 tctgctcatt gaccagcagg gctacattca tgtgacagac ttcggtttcg ccaagcgcgt
     1681 gaagggccgc acttggacct tgtgcggcac ccctgagtac ctggcccctg agattatcct
     1741 gagcaaaggc tacaacaagg ccgtggactg gtgggccctg ggggttctta tctatgaaat
     1801 ggccgctggc tacccgccct tcttcgcaga ccagcccatc cagatcctat atgagaaaat
     1861 cgtctctggg aaggtgcggt tcccatccca cttcagctct gacttgaagg acctgctgcg
     1921 gaaccttctg caggtagatc tcaccaagcg ctttgggaac ctcaagaatg gggtcaacga
     1981 tatcaagaac cacaagtggt ttgccacaac tgactggatt gccatctacc agaggaaggt
     2041 ggaagctccc ttcataccaa agtttaaagg ccctggggat acgagtaact ttgacgacta
     2101 tgaggaagaa gaaatccggg tctccatcaa tgagaagtgt ggcaaggagt tttctgagtt
     2161 tggatccggg agcatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     2221 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     2281 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     2341 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     2401 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     2461 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     2521 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     2581 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     2641 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     2701 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     2761 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     2821 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     2881 gtaatctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     2941 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3001 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3061 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3121 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3181 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3241 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3301 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3361 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3421 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3481 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3541 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3601 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3661 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3721 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3781 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3841 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3901 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3961 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4021 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4081 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4141 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4201 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4261 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4321 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4381 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4441 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4501 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4561 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4621 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4681 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4741 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4801 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4861 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4921 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4981 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5041 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5101 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5161 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5221 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5281 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5341 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5401 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5461 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5521 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5581 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5641 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5701 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5761 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5821 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5881 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5941 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6001 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6061 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     6121 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6181 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6241 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6301 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6361 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6421 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6481 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6541 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6601 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6661 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6721 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6781 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6841 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6901 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6961 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7021 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7081 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7141 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7201 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7261 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7321 tc
//
