LOCUS       sequence-490030-        5561 bp    DNA     circular SYN 21-OCT-2025
DEFINITION  Expressing constitutively active XAC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5561)
  AUTHORS   Dawe HR, Minamide LS, Bamburg JR, Cramer LP
  TITLE     ADF/cofilin controls cell polarity during fibroblast migration.
  JOURNAL   Curr Biol. 2003 Feb 4;13(3):252-7. doi: 
            10.1016/s0960-9822(03)00040-x.
  PUBMED    12573223
REFERENCE   2  (bases 1 to 5561)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 21, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Curr Biol"; date: "2003-02-4- 4"; volume: "13"; issue: "3"; pages: 
            "252-7"
COMMENT     Sequence Label: XAC1 S3A
FEATURES             Location/Qualifiers
     source          1..5561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     80..101
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        250..629
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        630..833
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     784..804
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     878..897
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        878..896
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      956..965
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1548..1565)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1554..1778
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(1910..2498)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1990..2009)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2827..2874
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2899..2918
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3106..3900)
                     /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             complement(3106..3900)
                     /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(3217..3236)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3827..3846
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3935..4292)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3949..4084
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4101..4292
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4249..4269
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4352..5143,5144..5212))
                     /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     4975..4994
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5213..5317)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5344..5561,1..238)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5431..5450)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
       61 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
      121 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
      181 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttac
      241 gcgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      301 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      361 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      421 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      481 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      541 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      601 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      661 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      721 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      781 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      841 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      901 cccaagctgg ctagcgttta aacttaagct tggtaccgag ctcggatccg cggccgccac
      961 catggccgct ggtgtgatgg tctccgatga tgtcattaaa gtatttaatg aaatgaaggt
     1021 gcgccatcaa ctctccccag aagacgccaa aaaacgcaag aaggcagttg tcttctgtct
     1081 tagcgacgac aagaaaacca tcattttgga accaggaaag gagatcttgc aaggagatat
     1141 cggttgtaac gttgaagatc cctacaagac ctttgtgaag atgctgcccc ggaatgactg
     1201 ccgttatgca ctgtatgatg cactctacga gacaaaggaa acaaagaaag aggatcttgt
     1261 ttttgtgttc tgggcccctg aagaagcctc acttaaaagt aaaatgatct acgctagctc
     1321 aaaagatgca atcaagaaga gattaccagg tatcaaacac gagtggcaaa taaacaccta
     1381 tgaagatgta aacgatccct gtaaccttgc ggataaacta gggggcaaca ctgtagtttc
     1441 ccttgaaggc aaatccgtga gaagttgaga tctcatcgaa ttctgcagat atccagcaca
     1501 gtggcggccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     1561 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     1621 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     1681 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     1741 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     1801 tggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1861 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1921 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1981 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2041 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2101 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2161 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2221 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2281 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2341 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2401 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2461 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2521 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2581 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2641 gtatatatga gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc
     2701 ctgggccttc acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt
     2761 ggccccaatg gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc
     2821 gtttatgaac aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag
     2881 cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg
     2941 cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa
     3001 cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca
     3061 ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc
     3121 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
     3181 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
     3241 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
     3301 attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc
     3361 gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc
     3421 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat
     3481 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg
     3541 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc
     3601 ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag
     3661 cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg
     3721 cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc
     3781 tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc
     3841 gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat
     3901 gcgaaacgat cctcatcctg tctcttgatc gatctttgca aaagcctagg cctccaaaaa
     3961 agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa
     4021 taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg
     4081 ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca
     4141 tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac taattgagat
     4201 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac
     4261 acacattcca cagctggttc tttccgcctc aggactcttc ctttttcaat aaatcaatct
     4321 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4381 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4441 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4501 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4561 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4621 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4681 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4741 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4801 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4861 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4921 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4981 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5041 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5101 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5161 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5221 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5281 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5341 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     5401 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     5461 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     5521 ttagtgcttt acggcacctc gaccccaaaa aacttgatta g
//
