LOCUS       Exported                7178 bp ds-DNA     circular SYN 04-MAY-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7178)
  AUTHORS   Adesnik Lab
  TITLE     Direct Submission
  JOURNAL   Exported May 4, 2021 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7178
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        386..661
                     /label=chicken beta-actin promoter
                     /note="color: #ffffff; direction: RIGHT"
     intron          662..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
                     /note="color: #------"
     CDS             1741..1758
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     CDS             1762..1794
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /note="color: #cc99b2"
                     /translation="MASMTGGQQMG"
     CDS             1798..1821
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /note="This feature has 2 segments:
                        1: 1798 .. 1806 / #cc99b2
                        2: 1807 .. 1821 / #cc99b2 / enterokinase site
                     Cleavage site after base 1821"
                     /translation="DLYDDDDK"
     misc_feature    1822..3081
                     /label=GCaMP6s
                     /note="color: #99cc00; direction: RIGHT"
     intron          3101..3234
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
                     /note="color: #------"
     misc_feature    3243..3831
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     misc_feature    3269..3288
                     /label=seq primer 5
                     /note="color: #a6acb3"
     CDS             complement(3714..3725)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 3713"
                     /translation="IEGR"
     polyA_signal    3864..3975
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     polyA_site      4267..4322
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(4681..4697)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    4705..4721
                     /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(4729..4759)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 4729 .. 4735 / #ffffff / -10
                        2: 4736 .. 4753 / #ffffff
                        3: 4754 .. 4759 / #ffffff / -35"
     protein_bind    4774..4795
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     promoter        4854..5049
                     /label=SV40 promoter
                     /note="SV40 early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      4900..5035
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     polyA_signal    5055..5189
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      complement(5428..6016)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6187..7047)
                     /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: 6187 .. 6978 / #ccffcc
                        2: 6979 .. 7047 / #ccffcc / signal sequence"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    6228..6247
                     /label=Seq primer 8
                     /note="color: #a6acb3"
     misc_feature    6875..6894
                     /label=Seq primer 7
                     /note="color: #a6acb3"
     promoter        complement(7048..7152)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg ggcgagggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcgg taccgccacc
     1741 catcatcatc atcatcatgg tatggctagc atgactggtg gacagcaaat gggtcgggat
     1801 ctgtacgacg atgacgataa ggatctcgcc accatggtcg actcatcacg tcgtaagtgg
     1861 aataagacag gtcacgcagt cagagctata ggtcggctga gctcactcga gaacgtctat
     1921 atcaaggccg acaagcagaa gaacggcatc aaggcgaact tccacatccg ccacaacatc
     1981 gaggacggcg gcgtgcagct cgcctaccac taccagcaga acacccccat cggcgacggc
     2041 cccgtgctgc tgcccgacaa ccactacctg agcgtgcagt ccaaactttc gaaagacccc
     2101 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     2161 ggcatggacg agctgtacaa gggcggtacc ggagggagca tggtgagcaa gggcgaggag
     2221 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     2281 ttcagcgtgt ccggcgaggg tgagggcgat gccacctacg gcaagctgac cctgaagttc
     2341 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     2401 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     2461 gccatgcccg aaggctacat ccaggagcgc accatcttct tcaaggacga cggcaactac
     2521 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     2581 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caacctgccg
     2641 gaccaactga ctgaagagca gatcgcagaa tttaaagagg aattctccct atttgacaag
     2701 gacggggatg ggacaataac aaccaaggag ctggggacgg tgatgcggtc tctggggcag
     2761 aaccccacag aagcagagct gcaggacatg atcaatgaag tagatgccga cggtgacggc
     2821 acaatcgact tccctgagtt cctgacaatg atggcaagaa aaatgaaata cagggacacg
     2881 gaagaagaaa ttagagaagc gttcggtgtg tttgataagg atggcaatgg ctacatcagt
     2941 gcagcagagc ttcgccacgt gatgacaaac cttggagaga agttaacaga tgaagaggtt
     3001 gatgaaatga tcagggaagc agacatcgat ggggatggtc aggtaaacta cgaagagttt
     3061 gtacaaatga tgacagcgaa gtaataactc gaggccgcag gtaagtatca aggttacaag
     3121 acaggtttaa ggagaccaca tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt
     3181 tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcg
     3241 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     3301 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     3361 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     3421 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     3481 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     3541 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     3601 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     3661 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     3721 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     3781 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaattc
     3841 gagctcggta cgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3901 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3961 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     4021 gcaggacagc aagggggagg attgggaaga caatagcagg ccagcttttg ttccctttag
     4081 ggttaattgc gcgcttggcg ctaatcatgg tcatagctgt ttccactcct caggtgcagg
     4141 ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag
     4201 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     4261 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     4321 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
     4381 agagtttggc aacatatgcc atatgctggc tgccatgaac aaaggtggct ataaagaggt
     4441 catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact
     4501 tgaggttaga ttttttttta tattttgttt gtgttatttt tttctttaac atccctaaaa
     4561 ttttccttac atgttttact agccagattt ttcctcctct cctgactact cccagtcata
     4621 gctgtccctc ttctcttatg aagatccctc gacctgcagc ccaagcttgg cgtaatcatg
     4681 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4741 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4801 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgcatctc
     4861 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4921 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4981 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5041 ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5101 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5161 tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg
     5221 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5281 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5341 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5401 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5461 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5521 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5581 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     5641 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5701 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5761 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5821 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     5881 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5941 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6001 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6061 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6121 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6181 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6241 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6301 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6361 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6421 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6481 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6541 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6601 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6661 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6721 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6781 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6841 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6901 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6961 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7021 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7081 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7141 aggggttccg cgcacatttc cccgaaaagt gccacctg
//
