LOCUS       sequence-229285-        8835 bp    DNA     circular SYN 14-APR-2025
DEFINITION  To express CARMIL2 with GFP tag on N-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8835)
  AUTHORS   Lanier MH, Kim T, Cooper JA
  TITLE     CARMIL2 is a novel molecular connection between vimentin and actin 
            essential for cell migration and invadopodia formation.
  JOURNAL   Mol Biol Cell. 2015 Dec 15;26(25):4577-88. doi: 
            10.1091/mbc.E15-08-0552. Epub 2015 Oct 14.
  PUBMED    26466680
REFERENCE   2  (bases 1 to 8835)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1091/mbc.E15-08-0552"; journalName: "Mol Biol Cell"; date: 
            "2015-12-15- 15"; volume: "26"; issue: "25"; pages: "4577-88"
COMMENT     Sequence Label: GFP-C-CARMIL2b
FEATURES             Location/Qualifiers
     source          1..8835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        118..421
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        422..625
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     576..596
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4839..5558
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    5680..5801
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5717..5736)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5771..5790
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5808..6263)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5945..5966)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6157..6176
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6290..6394
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6396..6753
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6396..6587
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6419..6439)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6604..6739
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6788..7582
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6788..7582
                     /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))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6842..6861)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7452..7471
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7770..7789)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7814..7861
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8190..8778
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8679..8698
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
       61 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      121 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      181 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      241 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      301 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      361 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      421 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      481 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      541 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      601 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg
      661 gactcagatc tatggcccag acccccgacg gcatctcctg tgagctccga ggcgagatca
      721 ccaggttcct gtggcccaaa gaggtggagc tgctgctgaa aacctggcta cccggggagg
      781 gtgctgtgca aaaccatgtc ctggcactgc tacgatggag agcctacctg ctgcacacca
      841 cctgcctccc gctgagggtg gactgcacgt tcagctacct ggaggtccag gccatggcgc
      901 tgcaggagac accccctcag gtcacctttg agctggagtc cctgcgtgag ctggtcctgg
      961 agtttcctgg tgtggccgcc ctggaacagc tggcccagca cgtggctgca gccatcaaga
     1021 aggtcttccc tcgctcgacc cttgggaagc tattccggag gcccacacca gcctccatgc
     1081 tggctcggct ggagagaagc agcccctcgg agtccactga cccctgcagc ccctgtggtg
     1141 gcttcttgga gacatacgag gctctgtgtg actacaatgg cttccctttc cgagaggaga
     1201 ttcagtggga cgtggacacc atttaccatc gccagggctg ccgccatttc agcctgggag
     1261 acttcagcca cctcggcagt cgggacctgg ccttgagtgt ggctgccctg tcctacaacc
     1321 tgtggttccg gtgcctctcc tgtgtggaca tgaagctgag ccttgaggtc tcagaacaga
     1381 ttctgcacat gatgagtcag tcatcacacc tggaggagct ggtgctggag acctgcagcc
     1441 tgaggggaga ctttgtccga cgactggccc aggcgctggc gggacactca agctctgggc
     1501 tgcgggagct cagcctcgcg gggaacctgc tggatgaccg aggcatgact gcactcagca
     1561 gacacctcga gcgttgtcca ggagccctga ggagactcag cctggcccag acagggttga
     1621 caccgcgagg aatgagggct ctgggccggg cactggccac caatgccgcc ttcgactcca
     1681 ccctgaccca cctggacctt tctgggaatc ctggggcgct gggggcctcc gaggacagtg
     1741 ggggcctcta tagcttcctg agccgtccta acgtactgtc gttcctgaat ctcgcaggca
     1801 ccgacactgc cctggacact ctcttcgcag cggtatcccg aggctgctgc accagcctta
     1861 cccacctcga cgcttcgagg aacgtcttct cccgcacgaa gtcccgcgcc gcgccggccg
     1921 cgctgcagct cttcctcagc cgcgcgcgga cgctgcggca cctgggcctg gcgggctgca
     1981 agctgccgcc cgacgcgctc agagcccttt tggatggcct cgcgctcaac acgcacctcc
     2041 gcgacctgca cctggacctc agcgcttgcg agctgcgctc ggccggcgcc caggtgatac
     2101 aagacttagt gtgcgacgca ggcgctgtga gctccctgga tctggcggat aacggcttcg
     2161 gctcagacat ggtgactctg gtgctggcca tcgggagaag ccggtccctg agacatgtgg
     2221 cgcttggaag gaacttcaac gtccggtgca aggagaccct ggacgacgtc ctgcaccgga
     2281 ttgtccagct catgcaggac gacgattgtc ctcttcagtc tctgtcggtg gctgagtctc
     2341 ggctgaagct gggtgccagc gtcctactcc gggccctagc caccaatcct aacctgaccg
     2401 cgctggatat cagcggcaac gccatggggg acgcgggcgc caagttgctg gccaaggcgc
     2461 tgcgggtcaa ctcgaggctc cggtctgtgg tctgggaccg gaaccacaca tctgctttgg
     2521 gtctgctgga cgtggcgcag gcgctggagc agaaccacag cctgaaggcc atgcctctgc
     2581 cactgaacga cgtggcccag gcgcagcgca gccgcccgga actgacagca cgtgcagtcc
     2641 atcagatcca agcctgtctc ttgaggaaca accgcgcaga ccctgcctct tctgaccaca
     2701 cgacccgcct tcagccactt ggtctggtct cagacccctc agagcaggaa gtgaatgaat
     2761 tgtgtcagtc ggtgcaggag catgtggagc tgctgggctg tggggctggg ccccagggtg
     2821 aagccgctgt gcgccaggcc gaggatgcca tccaaaatgc caacttctct ctcagcattc
     2881 tccccattct atatgaagct ggaagctccc caagccatca ctggcagctt gggcagaagc
     2941 tggagggcct tctgagacag gtgggcgagg tctgccgcca ggacatccag gacttcactc
     3001 aggccacact ggacacagca aggagcctct gcccacagat gctgcaggga tccagctgga
     3061 gggagcagct agagggggtc ctggcaggct cgaggggcct cccggagctg ctcccagagc
     3121 agctgctgca agatgccttc actaggctca gggacatgcg gctatcaatc acggggacct
     3181 tggcagagag cattgtggct caggctttgg caggcctgag tgcagcccgg gatcagctgg
     3241 tggagagtct ggctcagcag gcaacagtga caatgccccc tgccctacca gcaccggatg
     3301 gaggtgagcc cagcctcctt gagcctgggg aattggaagg tcttttcttc cccgaggaga
     3361 aggaagagga gaaggagaag gatgacagtc ctccacagaa atggcctgag ctcagccacg
     3421 gtcttcacct ggtccccttc attcacagtg ctgctgagga agcggagccg gagcccgagc
     3481 tggcggctcc gggagaagat gcagagccgc aggcggggcc gtccgcgcgc ggctctccga
     3541 gccctgccgc ccctgggccc ccggccggcc cgctgccccg catggacctg ccactggcgg
     3601 ggcagcccct gcgccatccg acccgggccc ggccgcggcc gcgccgccag caccaccacc
     3661 gcccgccgcc ggggggcccc caggtacccc cagccttgcc gcaggaaggg aatgggctca
     3721 gtgcccgcgt ggacgagggc gtggaggaat tcttctccaa aaggctgatc cagcaggatc
     3781 gcctctgggc ccccgaggag gacccggcca ctgagggggg cgccactcct gtcccccgta
     3841 cactgcgaaa gaagctgggc accctctttg ccttcaagaa gcctcgttca acgcggggtc
     3901 cacggactga tctagagacc agccctgggg cagctccccg aacccgaaaa actacatttg
     3961 gcgacctact gcggccgcca acccgtccca gccgtggtga ggagcttggt ggggctgagg
     4021 gggacaccag cagccctgac cctgccggca ggagccgacc tcgctacaca agagatagca
     4081 aggcctactc gatgatactg ctgcctgccg aggaggaggc aacgctgggt gccagacccg
     4141 acaagcggcg gcccctggag cggggagaaa cagaactggc tccatccttt gaacagcggg
     4201 tacaagtaat gctgcagagg ataggcgtca gccgaggcag cgggggtgcc gaaggcaaga
     4261 ggaagcaaag caaagatggc gagatcaaga aagctggctc agatggtgac attatggaca
     4321 gttccacgga ggcccctccc atctcgatca agtcccgcac ccactctgtg tctgctgacc
     4381 cttcctgcag acctggccca gggagccagg ggcctgagtc tgccacctgg aagacactgg
     4441 ggcagcagtt gaatgcggag ctcaggagcc gtggttgggg ccaacaggat ggtccaggcc
     4501 ctccctcccc tggtcaaagc ccaagtccct gcagaaccag cccctcccca gacagcctgg
     4561 gcctcccaga ggacccttgc ttgggcccca gaaatgaaga acggcccctg aggctgcagc
     4621 gctcccccgt cctcaaacgc aggccaaaac tcgaggcacc tccatcccca agcctaggat
     4681 ctggccttgg aaccgagcct ctgcccccac agcccacaga gccctccagc cctgagcgga
     4741 gcccaccctc cccagccaca gaccaaagag gcggcggccc caatcccaag cttcgaattc
     4801 tgcagtcgac ggtaccgcgg gcccgggatc caccggtcat ggtgagcaag ggcgccgagc
     4861 tgttcaccgg catcgtgccc atcctgatcg agctgaatgg cgatgtgaat ggccacaagt
     4921 tcagcgtgag cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     4981 tctgcaccac cggcaagctg cctgtgccct ggcccaccct ggtgaccacc ctgagctacg
     5041 gcgtgcagtg cttctcacgc taccccgatc acatgaagca gcacgacttc ttcaagagcg
     5101 ccatgcctga gggctacatc caggagcgca ccatcttctt cgaggatgac ggcaactaca
     5161 agtcgcgcgc cgaggtgaag ttcgagggcg ataccctggt gaatcgcatc gagctgaccg
     5221 gcaccgattt caaggaggat ggcaacatcc tgggcaataa gatggagtac aactacaacg
     5281 cccacaatgt gtacatcatg accgacaagg ccaagaatgg catcaaggtg aacttcaaga
     5341 tccgccacaa catcgaggat ggcagcgtgc agctggccga ccactaccag cagaataccc
     5401 ccatcggcga tggccctgtg ctgctgcccg ataaccacta cctgtccacc cagagcgccc
     5461 tgtccaagga ccccaacgag aagcgcgatc acatgatcta cttcggcttc gtgaccgccg
     5521 ccgccatcac ccacggcatg gatgagctgt acaagtgagc ggccgcgact ctagatcata
     5581 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     5641 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     5701 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5761 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     5821 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     5881 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     5941 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     6001 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     6061 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     6121 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     6181 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     6241 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     6301 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     6361 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     6421 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     6481 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     6541 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     6601 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     6661 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     6721 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     6781 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     6841 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     6901 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     6961 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     7021 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     7081 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     7141 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     7201 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     7261 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     7321 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     7381 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     7441 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     7501 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     7561 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     7621 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     7681 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     7741 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     7801 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     7861 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     7921 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     7981 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     8041 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     8101 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     8161 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     8221 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     8281 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     8341 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     8401 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     8461 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     8521 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     8581 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     8641 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     8701 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     8761 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     8821 tggccttttg ctcac
//
