LOCUS       Exported                6771 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    MyD116.PFLAG.CMV2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6771)
  AUTHORS   Novoa I, Zeng H, Harding HP, Ron D
  TITLE     Feedback inhibition of the unfolded protein response by 
            GADD34-mediated dephosphorylation of eIF2alpha.
  JOURNAL   J Cell Biol. 2001 May 28. 153(5):1011-22.
  PUBMED    11381086
REFERENCE   2  (bases 1 to 6771)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6771
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             935..958
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3112..3734
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3311..3330)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3758..3778)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3763..4092
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3943..4078
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4005..4024
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(4130..4149)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4131..4149)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4163..4179)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4163..4179)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4176..4198)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4187..4203
                     /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(4211..4241)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4256..4277
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4394..4411)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4565..5153)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4645..4664)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5324..6184)
                     /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"
     primer_bind     5947..5966
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6185..6289)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6316..6771
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6403..6422)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6613..6634
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
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 attgatctac catggactac aaagacgatg acgacaagct
      961 tgcggccggg gtgacgtgca gcccgccgcc cagacacatg gccccgagcc caagacccca
     1021 gcatgtcctg cactggaggg acgcccacaa cttctatctc ctgtccccac tgatgggctt
     1081 gctcagtcgg gcctggagcc gcctgagggg cccagaagtc ccagaggcat ggctggcaaa
     1141 aacagtaaca ggagcagatc agatagaagc tgcggctctg ctgacaccta cccctgtctc
     1201 tggtaacctc ctccctcatg gggagactga agaaagtgga tctcctgaac agagtcaagc
     1261 agcccagagg ctctgccttg tggaagctga aagttcccct cctgaaactt ggggactttc
     1321 aaatgttgat gagtacaatg caaagccagg acaagatgac cttagagaga aggaaatgga
     1381 acgcacagct ggcaaggcca cactacagcc cgctggcctg caaggggctg ataagaggct
     1441 tggggaggtg gtggctagag aagagggagt ggctgagccc gcttatccca catcacagct
     1501 ggagggtggt ccagctgaga atgaagagga tggagaaaca gtgaagactt accaagcttc
     1561 tgctgcttcc atagctccgg gatacaaacc cagcacccct gtgcctttct tgggggaggc
     1621 agaacatcaa gccacggaag aaaaaggaac agaaaacaag gctgacccct ccaactctcc
     1681 ttcttcaggc tcccactcca gagcctggga gtactactct agagagaagc ctaagcagga
     1741 gggagaagcc aaggtagagg cacacagggc agggcagggt cacccttgtc ggaatgctga
     1801 ggctgaggaa ggaggacctg agacaacttt tgtctgtact ggaaatgcct tcctgaaggc
     1861 ctgggtgtat cgcccaggag aggacacaga ggaagaagac aacagcgatt cggattcagc
     1921 tgaggaagac acagctcaga ccggtgccac cccccataca agtgccttcc tgaaggcctg
     1981 ggtgtatcgc ccaggagagg acacagagga agaagacagc gattcggatt cagctgagga
     2041 agacacagct cagaccggtg ccacccccca tacaagtgcc ttcctgaagg cctgggtgta
     2101 tcgcccagga gaggacacag aggaagaaaa cagcgatttg gattcagctg aggaagacac
     2161 agctcagacc ggtgccaccc cccatacaag tgccttcctg aaggcctggg tgtatcgccc
     2221 aggagaggac acagaggaag aaaacagcga tttggattca gctgaggaag acacagctca
     2281 gaccggtgcc accccacata caagtccctt cctgaaggcc tgggtgtatc gcccaggaga
     2341 ggacacagaa gatgacacag aagaggaaga ggacagtgag aatgtggccc caggtgactc
     2401 agaaacagct gactcaagcc agagtccctg ccttcagccc cagcgttgtc taccaggaga
     2461 gaagaccaag ggacgtgggg aagagccccc tctcttccag gtggccttct atttacccgg
     2521 agagaagcca gaatcacctt gggctgcacc taagctgccc cttcgactgc agaggcggct
     2581 cagattgttc aaagccccca cccgggatca ggaccccgag attcctctaa aagctcggaa
     2641 ggtacacttc gctgagaaag tcacagtcca tttccttgct gtctgggcag gaccagccca
     2701 agctgcccgt cgaggtccct gggagcagtt tgcacgagat cgaagccgct ttgctcgacg
     2761 cattgcccag gcagaggaga agctgggtcc ctaccttacc cctgattcca gggccagagc
     2821 atgggcacgc cttagaaacc catctcttcc acagtccgag cctcgctctt cctctgaggc
     2881 cactcccttg acccaagatg tgaccacacc ctctcccctt cccagtgaaa ccccttcgcc
     2941 cagcctgtac ttgggaggga ggcggggcta agcctgagta gtttcctatt atttatttat
     3001 ttatttattt gaataagaaa taaagccttt taatttgtag tgaaaaaaaa aaaaaaaaaa
     3061 ggaattcatc gatagatctg atatcggtac cagtcgactc tagaggatcc cgggtggcat
     3121 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     3181 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     3241 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     3301 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     3361 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     3421 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     3481 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     3541 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttaaaa
     3601 taactatacc agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg
     3661 gtgggacatt tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag
     3721 atgcctgttg aattgggtac gcggccagct tggctgtgga atgtgtgtca gttagggtgt
     3781 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3841 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3901 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3961 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4021 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4081 ggcttttgca aaaagctcct cgaggaactg aaaaaccaga aagttaattc cctatagtga
     4141 gtcgtattaa attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4201 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4261 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4321 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4381 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4441 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4501 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4561 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4621 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4681 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4741 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4801 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4861 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4921 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4981 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5041 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5101 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5161 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5221 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5281 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5341 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5401 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5461 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5521 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5581 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5641 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5701 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5761 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5821 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5881 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5941 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6001 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6061 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6121 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6181 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6241 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6301 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     6361 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     6421 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     6481 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     6541 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     6601 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     6661 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     6721 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt t
//
