LOCUS       Exported                5607 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  F-actin marker. EBFP2 is fused to the calponin homology domain of 
            utrophin (UtrCH)..
ACCESSION   .
VERSION     .
KEYWORDS    EBFP2-UtrCH
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5607)
  AUTHORS   Zhang Y, Cao F, Zhou Y, Feng Z, Sit B, Krendel M, Yu CH
  TITLE     Tail domains of myosin-1e regulate phosphatidylinositol signaling 
            and F-actin polymerization at the ventral layer of podosomes.
  JOURNAL   Mol Biol Cell. 2019 Mar 1;30(5):622-635. doi: 
            10.1091/mbc.E18-06-0398. Epub 2019 Jan 2.
  PUBMED    30601698
REFERENCE   2  (bases 1 to 5607)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5607
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1063..1072
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1069..1785
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(1114..1135)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(2615..2632)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2637..2771
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2661..2680)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2715..2734
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2884..2904)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2884..2902)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2923..2939)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2923..2939)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2936..2958)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2947..2963
                     /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(2971..3001)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3016..3037
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3154..3171)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3325..3913)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3405..3424)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4084..4944)
                     /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     4707..4726
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4945..5049)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5075..5530)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5212..5233)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5424..5443
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat ccgccaccat ggtgagcaag
     1081 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1141 ggccacaagt tcagcgtgag gggcgagggc gagggcgatg ccaccaacgg caagctgacc
     1201 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1261 ctgagccacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     1321 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     1381 ggcacctaca agacccgcgc cgaggtgaag ttcgagggcg acaccctagt gaaccgcatc
     1441 gagctgaagg gcgtcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1501 aacttcaaca gccacaacat ctatatcatg gccgtcaagc agaagaacgg catcaaggtg
     1561 aacttcaaga tccgccacaa cgtggaggac ggcagcgtgc agctcgccga ccactaccag
     1621 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acagccacta cctgagcacc
     1681 cagtccgtgc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1741 cgcaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg aaccatggcc
     1801 aagtatggag aacatgaagc cagtcctgac aatgggcaga acgaattcag tgatatcatt
     1861 aagtccagat ctgatgaaca caatgacgta cagaagaaaa cctttaccaa atggataaat
     1921 gctcgatttt caaagagtgg gaaaccaccc atcaatgata tgttcacaga cctcaaagat
     1981 ggaaggaagc tattggatct tctagaaggc ctcacaggaa catcactgcc aaaggaacgt
     2041 ggttccacaa gggtacatgc cttaaataac gtcaacagag tgctgcaggt tttacatcag
     2101 aacaatgtgg aattagtgaa tataggggga accgacattg tggatggaaa tcacaaactg
     2161 actttggggt tactttggag catcattttg cactggcagg tgaaagatgt catgaaggat
     2221 gtcatgtcgg acctgcagca gacgaacagt gagaagatcc tgctcagctg ggtgcgtcag
     2281 accaccaggc cctacagcca agtcaacgtc ctcaacttca ccaccagctg gacagatgga
     2341 ctcgccttta atgctgtcct ccaccgacat aaacctgatc tcttcagctg ggataaagtt
     2401 gtcaaaatgt caccaattga gagacttgaa catgccttca gcaaggctca aacttatttg
     2461 ggaattgaaa agctgttaga tcctgaagat gttgccgttc ggcttcctga caagaaatcc
     2521 ataattatgt atttaacatc tttgtttgag gtgctacctc agcaagtcac catagactaa
     2581 ctcgagctca agcttcgaat tctgcagtct agaactatag tgagtcgtat tacgtagatc
     2641 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     2701 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     2761 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     2821 gggaggtttt ttaattcgcg gccgcggcgc caatgcattg ggcccggtac ccagcttttg
     2881 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     2941 gtgaaattgt tatccgctca caattccaca caacatacga gccgggagca taaagtgtaa
     3001 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3061 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3121 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3181 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3241 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3301 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3361 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3421 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3481 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3541 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3601 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3661 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3721 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3781 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3841 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3901 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3961 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4021 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4081 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4141 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4201 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4261 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4321 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4381 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4441 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4501 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4561 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4621 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4681 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4741 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4801 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4861 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4921 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4981 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5041 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     5101 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     5161 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     5221 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     5281 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     5341 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     5401 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     5461 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     5521 cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     5581 gcctcttcgc tattacgcca gtcgacc
//
