LOCUS       Exported                7304 bp DNA     circular SYN 29-OCT-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7304)
  AUTHORS   Sohum Mehta
  TITLE     Direct Submission
  JOURNAL   Exported Jan 12, 2022 from SnapGene 6.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        105..484
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        485..688
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        733..751
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             787..2007
                     /codon_start=1
                     /label=DnaJB1(H33Q)-PKAcat
                     /note="color: #919191"
                     /translation="MGKDYYQTLGLARGASDEEIKRAYRRQALRYQPDKNKEPGAEEKF
                     KEIAEAYDVLSDPRKREIFDRYGEEVKEFLAKAKEDFLKKWESPAQNTAHLDQFERIKT
                     LGTGSFGRVMLVKHKETGNHYAMKILDKQKVVKLKQIEHTLNEKRILQAVNFPFLVKLE
                     FSFKDNSNLYMVMEYVPGGEMFSHLRRIGRFSEPHARFYAAQIVLTFEYLHSLDLIYRD
                     LKPENLLIDQQGYIHVTDFGFAKRVKGRTWTLCGTPEYLAPEIILSKGYNKAVDWWALG
                     VLIYEMAAGYPPFFADQPIQILYEKIVSGKVRFPSHFSSDLKDLLRNLLQVDLTKRFGN
                     LKNGVNDIKNHKWFATTDWIAIYQRKVEAPFIPKFKGPGDTSNFDDYEEEEIRVSINEK
                     CGKEFSEF"
     CDS             787..996
                     /codon_start=1
                     /label=DnaJB1 (1-70; H33Q)
                     /note="color: #945200"
                     /translation="MGKDYYQTLGLARGASDEEIKRAYRRQALRYQPDKNKEPGAEEKF
                     KEIAEAYDVLSDPRKREIFDRYGEE"
     CDS             997..2007
                     /codon_start=1
                     /label=PKA-Calpha (16-351)
                     /note="color: #942193"
                     /translation="VKEFLAKAKEDFLKKWESPAQNTAHLDQFERIKTLGTGSFGRVML
                     VKHKETGNHYAMKILDKQKVVKLKQIEHTLNEKRILQAVNFPFLVKLEFSFKDNSNLYM
                     VMEYVPGGEMFSHLRRIGRFSEPHARFYAAQIVLTFEYLHSLDLIYRDLKPENLLIDQQ
                     GYIHVTDFGFAKRVKGRTWTLCGTPEYLAPEIILSKGYNKAVDWWALGVLIYEMAAGYP
                     PFFADQPIQILYEKIVSGKVRFPSHFSSDLKDLLRNLLQVDLTKRFGNLKNGVNDIKNH
                     KWFATTDWIAIYQRKVEAPFIPKFKGPGDTSNFDDYEEEEIRVSINEKCGKEFSEF"
     CDS             2020..2730
                     /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    2771..2995
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      3041..3469
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3483..3812
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      3663..3798
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             3879..4673
                     /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    4847..4968
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(5017..5033)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    5041..5057
                     /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(5065..5095)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 5065 .. 5071 / #ffffff / -10
                        2: 5072 .. 5089 / #ffffff
                        3: 5090 .. 5095 / #ffffff / -35"
     protein_bind    5110..5131
                     /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(5419..6007)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6178..7038)
                     /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: 6178 .. 6969 / #ccffcc
                        2: 6970 .. 7038 / #ccffcc / signal sequence
                     Cleavage site after base 6969"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7039..7143)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
       61 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
      121 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      181 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      241 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      301 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      361 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      421 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      481 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      541 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      601 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      661 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      721 gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc gtttaaactt
      781 aagcttatgg gtaaagacta ctaccagacg ttgggcctgg cccgcggcgc gtcggacgag
      841 gagatcaagc gggcctaccg ccgccaggcg ctgcgctacc agccggacaa gaacaaggag
      901 cccggcgccg aggagaagtt caaggagatc gctgaggcct acgacgtgct cagcgacccg
      961 cgcaagcgcg agatcttcga ccgctacggg gaggaagtga aagaattctt agccaaagcc
     1021 aaagaagatt ttcttaaaaa atgggaaagt cccgctcaga acacagccca cttggatcag
     1081 tttgaacgaa tcaagaccct cggcacgggc tccttcgggc gggtgatgct ggtgaaacac
     1141 aaggagaccg ggaaccacta tgccatgaag atcctcgaca aacagaaggt ggtgaaactg
     1201 aaacagatcg aacacaccct gaatgaaaag cgcatcctgc aagctgtcaa ctttccgttc
     1261 ctcgtcaaac tcgagttctc cttcaaggac aactcaaact tatacatggt catggagtac
     1321 gtgcccggcg gggagatgtt ctcacaccta cggcggatcg gaaggttcag tgagccccat
     1381 gcccgtttct acgcggccca gatcgtcctg acctttgagt atctgcactc gctggatctc
     1441 atctacaggg acctgaagcc ggagaatctg ctcattgacc agcagggcta cattcatgtg
     1501 acagacttcg gtttcgccaa gcgcgtgaag ggccgcactt ggaccttgtg cggcacccct
     1561 gagtacctgg cccctgagat tatcctgagc aaaggctaca acaaggccgt ggactggtgg
     1621 gccctggggg ttcttatcta tgaaatggcc gctggctacc cgcccttctt cgcagaccag
     1681 cccatccaga tcctatatga gaaaatcgtc tctgggaagg tgcggttccc atcccacttc
     1741 agctctgact tgaaggacct gctgcggaac cttctgcagg tagatctcac caagcgcttt
     1801 gggaacctca agaatggggt caacgatatc aagaaccaca agtggtttgc cacaactgac
     1861 tggattgcca tctaccagag gaaggtggaa gctcccttca taccaaagtt taaaggccct
     1921 ggggatacga gtaactttga cgactatgag gaagaagaaa tccgggtctc catcaatgag
     1981 aagtgtggca aggagttttc tgagtttgga tccgggagca tggtgagcaa gggcgaggag
     2041 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     2101 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     2161 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     2221 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     2281 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     2341 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     2401 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     2461 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     2521 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     2581 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     2641 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     2701 accggcggca tggacgagct gtacaagtaa tagtctagag ggcccgttta aacccgctga
     2761 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2821 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2881 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2941 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3001 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3061 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3121 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3181 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3241 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3301 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3361 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3421 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3481 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3541 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3601 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3661 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3721 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3781 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttat atatccattt
     3841 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3901 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3961 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4021 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4081 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4141 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4201 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4261 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4321 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4381 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4441 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4501 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4561 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4621 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4681 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4741 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4801 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4861 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4921 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4981 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5041 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5101 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5161 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5221 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5281 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5341 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5401 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5461 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5521 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5581 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5641 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5701 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5761 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5821 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5881 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5941 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6001 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6061 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6121 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6181 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6241 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6301 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6361 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6421 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6481 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6541 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6601 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6661 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6721 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6781 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6841 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6901 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6961 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7021 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7081 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7141 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     7201 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     7261 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaat
//
