LOCUS       Exported                6610 bp ds-DNA     circular SYN 16-MAR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6610)
  AUTHORS   SnapGene License
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 20, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6610
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1493..3323
                     /organism="Homo sapiens"
                     /chromosome="X"
                     /map="Xq13.3"
                     /mol_type="mRNA"
                     /db_xref="taxon:9606"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     primer_bind     141..190
                     /label=F-D1
     primer_bind     147..194
                     /label=F-Mlu-D2
     primer_bind     147..161
                     /label=3'-pcalm-ampF
     primer_bind     150..168
                     /label=P-Calm1-F
     primer_bind     151..200
                     /label=F-D2
     misc_feature    156..276
                     /label=P Calm1
     misc_feature    156..165
                     /label=D1
     primer_bind     161..210
                     /label=F-D3
     primer_bind     complement(161..185)
                     /label=R-D4
     misc_feature    166..175
                     /label=D2
     primer_bind     171..220
                     /label=F-D4
     primer_bind     complement(171..195)
                     /label=R-D5
     misc_feature    176..185
                     /label=D3
     primer_bind     181..230
                     /label=F-D5
     primer_bind     complement(181..205)
                     /label=R-D6
     misc_feature    186..195
                     /label=D4
     primer_bind     191..240
                     /label=F-D6
     primer_bind     complement(191..215)
                     /label=r-d7
     misc_feature    196..205
                     /label=D5
     primer_bind     200..250
                     /label=F-D7
     primer_bind     complement(201..225)
                     /label=R-D8
     misc_feature    206..215
                     /label=D6
     primer_bind     211..260
                     /label=F-D8
     primer_bind     complement(215..236)
                     /label=R-D9
     misc_feature    216..225
                     /label=D7
     primer_bind     222..271
                     /label=F-D9
     primer_bind     complement(222..246)
                     /label=R-D10
     misc_feature    226..235
                     /label=D8
     primer_bind     complement(232..256)
                     /label=R-D11
     misc_feature    236
                     /label=+1
     misc_feature    237..246
                     /label=D9
     primer_bind     complement(242..266)
                     /label=R-D12
     misc_feature    247..256
                     /label=D10
     misc_feature    257..266
                     /label=D11
     primer_bind     complement(260..287)
                     /label=2B-1R
     primer_bind     complement(261..276)
                     /label=P-Calm1-R
     misc_feature    267..276
                     /label=D12
     primer_bind     271..296
                     /label=2B-2F
     primer_bind     complement(277..298)
                     /label=Primer 1
     misc_feature    277..282
                     /label=BsiWI
     primer_bind     283..302
                     /label=Primer 5
     CDS             288..1318
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     misc_feature    548
                     /label=silent mut
     primer_bind     complement(616..650)
                     /label=R-Cre-R119STOP
     primer_bind     complement(632..651)
                     /label=R-CRE-L115-BamHI
     primer_bind     636..670
                     /label=F-Cre-R119STOP
     primer_bind     1093..1116
                     /label=F-Seq-Cre-3F1
     misc_feature    1093..1098
                     /label=ClaI
     misc_feature    1224
                     /label=mut
     primer_bind     1262..1279
                     /label=cre-SF
     primer_bind     complement(1296..1318)
                     /label=R-T5-Cre
     intron          1320..1360
                     /label=Splicing donor
                     /note="Splicing donor"
     primer_bind     complement(1338..1367)
                     /label=2B-2R
     primer_bind     complement(1341..1385)
                     /label=Primer 7
     primer_bind     complement(1341..1385)
                     /label=Primer 6
     primer_bind     1350..1387
                     /label=2B-3F
     primer_bind     complement(1356..1380)
                     /label=Primer 3
     primer_bind     1367..1401
                     /label=Primer 8
     primer_bind     1367..1401
                     /label=Primer 2
     primer_bind     1367..1401
                     /label=F-T5-JT+2272
     protein_bind    1367..1400
                     /label=loxJTZ17?2272?
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    1380..1387
                     /label=2272
     intron          1401..1492
                     /label=Splicing acceptor
                     /note="Splicing acceptor"
     primer_bind     1401..1420
                     /label=Primer 4
     primer_bind     1459..1478
                     /label=T0060_PIRES-F
     primer_bind     complement(1468..1492)
                     /label=#57-R-SA-3R1
     primer_bind     1477..1512
                     /label=A14F-NheI-F
     primer_bind     1477..1492
                     /label=Primer 10
     misc_feature    1492
                     /label=NheI?
     CDS             1493..3323
                     /codon_start=1
                     /gene="NEXMIF"
                     /gene_synonym="KIAA2022; KIDLIA; MRX98; XLID98; XPN"
                     /product="neurite extension and migration factor"
                     /label=NEXMIF
                     /note="XLMR protein related to neurite extension;
                     XLMR-related protein, neurite extension"
                     /db_xref="GI:56711286"
                     /db_xref="CCDS:CCDS35337.1"
                     /db_xref="GeneID:340533"
                     /db_xref="HGNC:HGNC:29433"
                     /db_xref="MIM:300524"
                     /protein_id="NP_001008537.1"
                     /translation="EIAPTQSSEFGASQVVSMENNLTPTTYNPICLNSGGSNCNKVLYD
                     SMQDTQLPSDDSYQLCHFNNGEICFPFQQGPVNMDDGRLFSFDSMAPLSVSSSNYCSLS
                     LKSCEKDGDDDITDDFLAHCSPKLVIQQSIDEIAPLKESTDLLDISNFTPDKFRHSSLS
                     EMSPPDTPSLSPQITRCESMKTLGTLKGFQEGVPGPLDSVEKIKWDCSTLSRQVQMEDG
                     FTLNNHQFQFHMFNDEDSVSLLQKNPCLSTFNDPSGQISTNNKVSKSRKKSSPSKSGAM
                     NQSSSQKNTRKKSLKGNNKGIEKPPGKNSRQVPKSTKKGKYMAAINGEKMQIGIGRGGS
                     QTNTISSTGKTLAECIQHGGPMASMKMPSQKGLSGDWALGKESSPGWSDMSMGTNTNSL
                     LDDDQREFQEPSYILSNIASGMADVQRFMMASIEPLWEPMEHHGDPNIFYSPESNSLKL
                     KTLKILAGTPQESKKKINSGSQGATKNHRSIKGVSKSNGKTAIGDPGRANMPGYNEDSR
                     STFFDKKYSNMSTLGNNGPTHKKLYRHKSSSKALRDEKCKGKHMEREQVHKDESGTASF
                     EKLRDSDYNLLKAETTFWVLPVFEEETRIFQKDI"
     misc_feature    1493..1494
                     /label=Feature 16
     primer_bind     complement(2629..2645)
                     /label=14B-NEXMIF-SR
     primer_bind     complement(3303..3344)
                     /label=A14F-BsrGI-R
     misc_feature    3324..3329
                     /label=BsrGI
     CDS             3330..3395
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3335..3423)
                     /label=296
     terminator      3399..3830
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     primer_bind     3399..3424
                     /label=#56-F-W3SL-5F1
     primer_bind     complement(3478..3501)
                     /label=R-Seq-W3SL-5R1
     misc_feature    3873..4013
                     /label=R-ITR
     rep_origin      4016..4471
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4163..4172)
                     /label=F-Seq-Cre-3F1
     promoter        4753..4857
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4858..5718
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="HSNMYPLMRQ*P**MLQ*Y*KRKSMSIQHFRVALIPFFAAFCLPV
                     FAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLS
                     RIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTT
                     IGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLT
                     LASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIV
                     VIYTTG"
     rep_origin      5889..6477
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtcgctc tgcgcgaggc ggctccgccg
      181 cggcggaggg atacggcgca ccatatatat atcgcggggc gcagactcgc gctccggcag
      241 tggtgctggg agtgtcgtgg acgccgtgcc gttactcgta cgccaccatg tccaatttac
      301 tgaccgtaca ccaaaatttg cctgcattac cggtcgatgc aacgagtgat gaggttcgca
      361 agaacctgat ggacatgttc agggatcgcc aggcgttttc tgagcatacc tggaaaatgc
      421 ttctgtccgt ttgccggtcg tgggcggcat ggtgcaagtt gaataaccgg aaatggtttc
      481 ccgcagaacc tgaagatgtt cgcgattatc ttctatatct tcaggcgcgc ggtctggcag
      541 taaaaaccat ccagcaacat ttgggccagc taaacatgct tcatcgtcgg tccgggctgc
      601 cacgaccaag tgacagcaat gctgtttcac tggttatgcg gcggatccga aaagaaaacg
      661 ttgatgccgg tgaacgtgca aaacaggctc tagcgttcga acgcactgat ttcgaccagg
      721 ttcgttcact catggaaaat agcgatcgct gccaggatat acgtaatctg gcatttctgg
      781 ggattgctta taacaccctg ttacgtatag ccgaaattgc caggatcagg gttaaagata
      841 tctcacgtac tgacggtggg agaatgttaa tccatattgg cagaacgaaa acgctggtta
      901 gcaccgcagg tgtagagaag gcacttagcc tgggggtaac taaactggtc gagcgatgga
      961 tttccgtctc tggtgtagct gatgatccga ataactacct gttttgccgg gtcagaaaaa
     1021 atggtgttgc cgcgccatct gccaccagcc agctatcaac tcgcgccctg gaagggattt
     1081 ttgaagcaac tcatcgattg atttacggcg ctaaggatga ctctggtcag agatacctgg
     1141 cctggtctgg acacagtgcc cgtgtcggag ccgcgcgaga tatggcccgc gctggagttt
     1201 caataccgga gatcatgcaa gctggtggct ggaccaatgt aaatattgta atgaactata
     1261 tccgtaacct ggatagtgaa acaggggcaa tggtgcgcct gctggaagat ggcgattagg
     1321 taagtatcaa ggttacaaga caggtttaag gagaccaata actagtataa cttcgtatag
     1381 gatactttat agcaatttat gaaactgggc ttgtcgagac agagaagact cttgcgtttc
     1441 tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac agggagatag
     1501 ccccaaccca atccagtgaa tttggagcct cccaagtagt ctccatggaa aataacctca
     1561 cacctacaac atacaatcca atctgtctca atagtggtgg cagtaattgc aacaaggtcc
     1621 tgtatgactc catgcaagat acccaactcc catctgatga ctcttaccaa ttatgtcact
     1681 ttaataatgg agagatctgc tttcctttcc agcaggggcc agtcaatatg gatgatggtc
     1741 ggctctttag ctttgattca atggccccac tctctgtcag ctcaagtaat tattgctcct
     1801 taagcttgaa gtcctgtgaa aaggatggcg atgatgatat cactgatgac ttcctggccc
     1861 attgcagccc taagctggtg atccagcaaa gcattgatga gatagcacca ctaaaggagt
     1921 ccactgacct cctggatata tccaacttca cccctgacaa attccgccac tcttcccttt
     1981 cagagatgtc cccaccggac acccctagtc tttcccctca aattaccaga tgtgagagta
     2041 tgaagacact aggaacacta aaggggttcc aagagggtgt cccaggacca ttggacagtg
     2101 tggaaaaaat caagtgggac tgcagtaccc tttcacggca ggtccaaatg gaggatggat
     2161 ttactttaaa taatcaccag tttcagttcc atatgttcaa tgatgaggat tctgtcagcc
     2221 tgctccaaaa aaacccttgc ctgtcaacat ttaatgatcc atctggtcaa attagtacca
     2281 acaacaaagt gtcaaaatca agaaagaaaa gttcacccag caagagtggg gctatgaacc
     2341 aaagctcttc tcagaaaaac accaggaaaa aatccctcaa aggtaacaac aaggggattg
     2401 aaaaaccacc tggcaaaaac tcccgccagg tccctaagtc cacaaagaaa gggaaataca
     2461 tggctgccat caatggagag aaaatgcaaa ttggcattgg ccgtggggga agccaaacca
     2521 acaccatatc ctccactgga aagacattgg ctgaatgtat ccaacatggt ggccctatgg
     2581 cctctatgaa gatgccaagt caaaagggac tttctggaga ttgggccttg gggaaggaga
     2641 gcagcccagg ctggagtgat atgagcatgg gcaccaacac caacagcctt ctggatgatg
     2701 accaacggga atttcaggag ccttcctata tcttgtccaa cattgcctct ggtatggcag
     2761 atgtgcagag attcatgatg gcctccatag agcccctttg ggaacccatg gagcaccatg
     2821 gggatcccaa catattctac tcccctgagt ccaatagtct aaaattaaaa accctcaaaa
     2881 tattggctgg gacaccacag gagtctaaga aaaagatcaa cagtgggtcc caaggagcca
     2941 ctaagaatca caggtcaatc aagggtgtga gcaaaagcaa tgggaaaaca gcaataggtg
     3001 atcctggtcg tgcaaacatg cctggttata acgaggactc tcgctctacc ttctttgata
     3061 aaaagtatag taacatgagc actttaggca ataacggacc gacacacaaa aagttgtatc
     3121 gtcacaaatc cagctccaag gccctgagag acgagaaatg taagggaaag cacatggagc
     3181 gagaacaggt ccacaaggat gagtctggga cagcttcttt tgaaaaactg agggattccg
     3241 actacaatct cctaaaagca gaaacaacct tttgggtttt acctgtgttt gaagaagaga
     3301 ctcgcatttt ccagaaagac atttgtacag actacaaaga ccatgacggt gattataaag
     3361 atcatgacat cgactacaag gatgacgatg acaagtaact cgagataatc aacctctgga
     3421 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3481 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3541 ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct
     3601 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgtttatttg
     3661 tgaaatttgt gatgctattg ctttatttgt aaccatctag ctttatttgt gaaatttgtg
     3721 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     3781 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa agatctggag
     3841 accggtgtgg cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     3901 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     3961 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     4021 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     4081 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4141 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     4201 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4261 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     4321 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4381 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     4441 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     4501 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     4561 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     4621 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4681 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     4741 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4801 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4861 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4921 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4981 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5041 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5101 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5161 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5221 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5281 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5341 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5401 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5461 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5521 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5581 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5641 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5701 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5761 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5821 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5881 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5941 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6001 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6061 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6121 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6181 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6241 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6301 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6361 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6421 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6481 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6541 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6601 gctcacatgt
//
