LOCUS       sequence-490239-        5561 bp    DNA     circular SYN 22-OCT-2025
DEFINITION  Expressing constitutively active XAC1 with lower turnover rate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5561)
  AUTHORS   Abe H, Obinata T, Minamide LS, Bamburg JR
  TITLE     Xenopus laevis actin-depolymerizing factor/cofilin: a 
            phosphorylation-regulated protein essential for development.
  JOURNAL   J Cell Biol. 1996 Mar;132(5):871-85. doi: 10.1083/jcb.132.5.871.
  PUBMED    8603919
REFERENCE   2  (bases 1 to 5561)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1083/jcb.132.5";
            journalName: "J Cell Biol"; date: "1996-03"; volume: "132"; issue: 
            "5"; pages: "871-85"
COMMENT     Sequence Label: XAC1 S3A,Y82F
FEATURES             Location/Qualifiers
     source          1..5561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(150..167)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    156..380
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(512..1100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(592..611)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1429..1476
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1501..1520
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1708..2502)
                     /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(1708..2502)
                     /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(1819..1838)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2429..2448
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2537..2894)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2551..2686
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        2703..2894
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2851..2871
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(2954..3745,3746..3814))
                     /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     3577..3596
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3815..3919)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3946..4401
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4033..4052)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4243..4264
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        4413..4792
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4793..4996
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4947..4967
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5041..5060
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5041..5059
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5119..5128
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctgtaacctt gcggataaac tagggggcaa cactgtagtt tcccttgaag gcaaatccgt
       61 gagaagttga gatctcatcg aattctgcag atatccagca cagtggcggc cgctcgagtc
      121 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
      181 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
      241 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
      301 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
      361 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agtggcggta atacggttat
      421 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
      481 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
      541 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
      601 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
      661 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
      721 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
      781 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
      841 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
      901 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
      961 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1021 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1081 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     1141 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     1201 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg
     1261 aggctatggc agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac
     1321 ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg
     1381 gtggggtatc gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc
     1441 caacaccgtg cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta
     1501 ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga
     1561 gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc
     1621 tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt
     1681 cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc
     1741 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc
     1801 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc
     1861 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt
     1921 cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt
     1981 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg
     2041 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg
     2101 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat
     2161 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc
     2221 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac
     2281 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc
     2341 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc
     2401 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca
     2461 agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc
     2521 tgtctcttga tcgatctttg caaaagccta ggcctccaaa aaagcctcct cactacttct
     2581 ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc
     2641 catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg
     2701 gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg
     2761 agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc
     2821 tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc cacagctggt
     2881 tctttccgcc tcaggactct tcctttttca ataaatcaat ctaaagtata tatgagtaaa
     2941 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3001 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3061 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3121 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3181 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3241 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3301 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3361 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3421 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3481 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3541 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     3601 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     3661 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     3721 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     3781 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     3841 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     3901 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg
     3961 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4021 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4081 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4141 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4201 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4261 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     4321 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4381 aaatattaac gcttacaatt tacgcgcgcg ttgacattga ttattgacta gttattaata
     4441 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     4501 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     4561 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     4621 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     4681 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     4741 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     4801 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     4861 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     4921 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     4981 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
     5041 taatacgact cactataggg agacccaagc tggctagcgt ttaaacttaa gcttggtacc
     5101 gagctcggat ccgcggccgc caccatggcc gctggtgtga tggtctccga tgatgtcatt
     5161 aaagtattta atgaaatgaa ggtgcgccat caactctccc cagaagacgc caaaaaacgc
     5221 aagaaggcag ttgtcttctg tcttagcgac gacaagaaaa ccatcatttt ggaaccagga
     5281 aaggagatct tgcaaggaga tatcggttgt aacgttgaag atccctacaa gacctttgtg
     5341 aagatgctgc cccggaatga ctgccgtttt gcactgtatg atgcactcta cgagacaaag
     5401 gaaacaaaga aagaggatct tgtttttgtg ttctgggccc ctgaagaagc ctcacttaaa
     5461 agtaaaatga tctacgctag ctcaaaagat gcaatcaaga agagattacc aggtatcaaa
     5521 cacgagtggc aaataaacac ctatgaagat gtaaacgatc c
//
