LOCUS       Exported                6807 bp ds-DNA     circular SYN 02-MAR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6807)
  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..6807
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1493..3520
                     /organism="Homo sapiens"
                     /chromosome="5"
                     /map="5p13.2"
                     /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     1475..1492
                     /label=new-NheI-2B-F
     primer_bind     1477..1513
                     /label=A3F-NheI-F
     primer_bind     1477..1492
                     /label=Primer 10
     misc_feature    1492
                     /label=NheI?
     CDS             1493..3520
                     /codon_start=1
                     /gene="NUP155"
                     /gene_synonym="ATFB15; N155"
                     /product="nuclear pore complex protein Nup155 isoform 1"
                     /label=NUP155
                     /note="isoform 1 is encoded by transcript variant 1;
                     nuclear pore complex protein Nup155; 155 kDa nucleoporin; 
                     nucleoporin 155kD; nucleoporin 155kDa"
                     /db_xref="GI:24430149"
                     /db_xref="CCDS:CCDS3921.1"
                     /db_xref="GeneID:9631"
                     /db_xref="HGNC:HGNC:8063"
                     /db_xref="MIM:606694"
                     /protein_id="NP_705618.1"
                     /translation="GLQEFLDRNSQFAGGPLGNPNTTAKVQQRLIGFMRPENGNPQQMQ
                     QELQRKFHEAQLSEKISLQAIQQLVRKSYQALALWKLLCEHQFTIIVAELQKELQEQLK
                     ITTFKDLVIRDKELTGALIASLINCYIRDNAAVDGISLHLQDICPLLYSTDDAICSKAN
                     ELLQRSRQVQNKTEKERMLRESLKEYQKISNQVDLSNVCAQYRQVRFYEGVVELSLTAA
                     EKKDPQGLGLHFYKHGEPEEDIVGLQAFQERLNSYKCITDTLQELVNQSKAAPQSPSVP
                     KKPGPPVLSSDPNMLSNEEAGHHFEQMLKLSQRSKDELFSIALYNWLIQVDLADKLLQV
                     ASPFLEPHLVRMAKVDQNRVRYMDLLWRYYEKNRSFSNAARVLSRLADMHSTEISLQQR
                     LEYIARAILSAKSSTAISSIAADGEFLHELEEKMEVARIQLQIQETLQRQYSHHSSVQD
                     AVSQLDSELMDITKLYGEFADPFKLAECKLAIIHCAGYSDPILVQTLWQDIIEKELSDS
                     VTLSSSDRMHALSLKIVLLGKIYAGTPRFFPLDFIVQFLEQQVCTLNWDVGFVIQTMNE
                     IGVPLPRLLEVYDQLFKSRDPFWNRMKKPLHLLDCIHVLLIRYVENPSQVLNCERRRFT
                     NLCLDAVCGYLVELQSMSSSVAVQAITGNFKSLQAKLERLH"
     misc_feature    1493..1494
                     /label=Feature 46
     exon            1653..1783
                     /gene="NUP155"
                     /gene_synonym="ATFB15; N155"
                     /inference="alignment:same species:2.1.0"
     primer_bind     2168..2184
                     /label=A3F-SF
     primer_bind     complement(2748..2764)
                     /label=3B-NUP155-SR
     primer_bind     complement(3497..3529)
                     /label=R1
     primer_bind     complement(3500..3541)
                     /label=nA3F-BsrGI-R
     primer_bind     complement(3500..3541)
                     /label=A3F-BsrGI-R
     primer_bind     complement(3500..3520)
                     /label=Primer 11
     primer_bind     3512..3552
                     /label=F2
     primer_bind     3521..3546
                     /label=Primer 9
     misc_feature    3521..3526
                     /label=BsrGI
     CDS             3527..3592
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3532..3620)
                     /label=296
     terminator      3596..4027
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     primer_bind     3596..3621
                     /label=#56-F-W3SL-5F1
     primer_bind     complement(3675..3698)
                     /label=R-Seq-W3SL-5R1
     misc_feature    4070..4210
                     /label=R-ITR
     rep_origin      4213..4668
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4360..4369)
                     /label=F-Seq-Cre-3F1
     promoter        4950..5054
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5055..5915
                     /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      6086..6674
                     /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 agggtttgca
     1501 ggaatttcta gacagaaact cccagtttgc aggaggacca ttaggaaatc caaatactac
     1561 tgctaaagtg cagcagaggc tgataggatt catgcgtcct gaaaacggaa atccccagca
     1621 aatgcaacag gaactgcaga ggaagtttca tgaggctcaa ctaagtgaaa agatttcact
     1681 tcaggcaatt cagcagttgg ttcgaaaatc atatcaggct ctggctttat ggaaacttct
     1741 ttgtgaacat caattcacta tcattgtggc agaacttcag aaggaacttc aagagcagct
     1801 gaagatcacc acctttaaag atcttgtaat cagggacaaa gaactcacag gggcattaat
     1861 tgcttctctt atcaactgct acatcagaga taatgccgct gttgatggca ttagtttaca
     1921 tttacaggat atctgcccac ttctatatag cactgatgat gcaatttgtt ctaaggcaaa
     1981 tgagcttctc cagcgttccc gacaagttca aaataagact gaaaaagaaa gaatgttaag
     2041 ggaatcatta aaggaatatc aaaaaattag caatcaagtg gacctttcca atgtttgtgc
     2101 tcagtataga caagtgagat tttatgaggg tgtggtggaa ctttctctta cggctgcaga
     2161 gaaaaaagat cctcaaggtc ttgggcttca tttctataaa catggagaac cagaagaaga
     2221 catagttgga cttcaggcct tccaagaaag attaaacagt tacaaatgca ttacagacac
     2281 acttcaagaa ctggtaaatc aaagtaaggc cgctcctcag tctcccagtg tacccaaaaa
     2341 acctggtcct ccagtgttgt catctgatcc aaatatgctg agtaatgaag aagcaggaca
     2401 tcattttgaa caaatgctta aattgtcaca gcgatccaag gatgagctct ttagtattgc
     2461 cctttataat tggctaatac aagtcgacct tgcagataag ctgctacagg ttgcttctcc
     2521 atttctggag ccacatctag tccgaatggc caaagttgat caaaacagag ttcgttatat
     2581 ggatttactc tggcggtatt acgagaagaa cagaagtttc agtaatgctg ctcgtgtact
     2641 gtccagactg gctgacatgc atagcacaga aatttcactt cagcagcgac tagagtacat
     2701 tgctcgagcc attcttagtg ccaaaagttc cactgccatt tcatcaatag ctgccgatgg
     2761 tgaatttctt catgaattag aagaaaaaat ggaggttgct aggatccaac ttcagataca
     2821 ggagacacta caaaggcagt attcccatca ttcttctgta caggatgcag tttctcagct
     2881 ggattctgag ctgatggaca taactaagct ttatggggaa tttgctgacc catttaaact
     2941 tgcagagtgc aaacttgcaa taattcattg tgccggttat tcagacccta tattggtgca
     3001 gacactttgg caagatatca tagagaaaga attgagtgac agtgtgacat tgagctcctc
     3061 ggatagaatg catgctctta gtctcaagat tgttctcctt ggcaaaattt atgctggcac
     3121 accacgcttc tttcctttag attttattgt acagttttta gaacagcagg tttgtacttt
     3181 gaactgggat gtgggcttcg taatacagac aatgaatgaa attggagtac cattacctag
     3241 actactagaa gtttatgatc agttgttcaa atcacgggat ccattctgga acagaatgaa
     3301 gaagccactg caccttttgg attgtataca tgtattattg ataagatatg ttgagaatcc
     3361 cagccaagtt ttaaattgtg aaaggagaag atttacaaat ctctgcctgg atgctgtttg
     3421 tggttacctt gttgagctcc agtctatgag ctcgtcagta gcagtacaag ccatcactgg
     3481 gaattttaaa tctcttcaag ctaaattaga acggcttcat tgtacagact acaaagacca
     3541 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtaactcga
     3601 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3661 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3721 cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa
     3781 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3841 tccgtggtgt ttatttgtga aatttgtgat gctattgctt tatttgtaac catctagctt
     3901 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     3961 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt
     4021 tttttaaaga tctggagacc ggtgtggcac gtgcggaccg agcggccgca ggaaccccta
     4081 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     4141 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     4201 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4261 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4321 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg
     4381 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4441 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4501 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4561 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta
     4621 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa
     4681 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     4741 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     4801 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     4861 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     4921 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     4981 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5041 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5101 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5161 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5221 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5281 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     5341 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     5401 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     5461 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5521 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     5581 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5641 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     5701 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5761 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5821 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     5881 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5941 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6001 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6061 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6121 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6181 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6241 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6301 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6361 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6421 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6481 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6541 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6601 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6661 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6721 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6781 ccttttgctg gccttttgct cacatgt
//
