LOCUS       Exported                7022 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 7022)
  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..7022
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1493..3735
                     /organism="Homo sapiens"
                     /chromosome="15"
                     /map="15q24.1"
                     /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..1532
                     /label=B21F-NheI-F
     primer_bind     1477..1492
                     /label=Primer 10
     misc_feature    1492
                     /label=NheI?
     CDS             1493..3735
                     /codon_start=1
                     /gene="NEO1"
                     /gene_synonym="IGDCC2; NGN; NTN1R2"
                     /product="neogenin isoform 1 precursor"
                     /label=NEO1
                     /note="isoform 1 precursor is encoded by transcript
                     variant 1; immunoglobulin superfamily DCC subclass member 
                     2; neogenin homolog 1"
                     /db_xref="GI:157311649"
                     /db_xref="CCDS:CCDS10247.1"
                     /db_xref="GeneID:4756"
                     /db_xref="HGNC:HGNC:7754"
                     /db_xref="MIM:601907"
                     /protein_id="NP_002490.2"
                     /translation="LDRGTEYNFRVAALTINGTGPATDWLSAETFESDLDETRVPEVPS
                     SLHVRPLVTSIVVSWTPPENQNIVVRGYAIGYGIGSPHAQTIKVDYKQRYYTIENLDPS
                     SHYVITLKAFNNVGEGIPLYESAVTRPHTVPDPTPMMPPVGVQASILSHDTIRITWADN
                     SLPKHQKITDSRYYTVRWKTNIPANTKYKNANATTLSYLVTGLKPNTLYEFSVMVTKGR
                     RSSTWSMTAHGTTFELVPTSPPKDVTVVSKEGKPKTIIVNWQPPSEANGKITGYIIYYS
                     TDVNAEIHDWVIEPVVGNRLTHQIQELTLDTPYYFKIQARNSKGMGPMSEAVQFRTPKA
                     DSSDKMPNDQASGSGGKGSRLPDLGSDYKPPMSGSNSPHGSPTSPLDSNMLLVIIVSVG
                     VITIVVVVIIAVFCTRRTTSHQKKKRAACKSVNGSHKYKGNSKDVKPPDLWIHHERLEL
                     KPIDKSPDPNPIMTDTPIPRNSQDITPVDNSMDSNIHQRRNSYRGHESEDSMSTLAGRR
                     GMRPKMMMPFDSQPPQPVISAHPIHSLDNPHHHFHSSSLASPARSHLYHPGSPWPIGTS
                     MSLSDRANSTESVRNTPSTDTMPASSSQTCCTDHQDPEGATSSSYLASSQEEDSGQSLP
                     TAHVRPSHPLKSFAVPAIPPPGPPTYDPALPSTPLLSQQALNHHIHSVKTASIGTLGRS
                     RPPMPVVVPSAPEVQETTRMLEDSESSYEPDELTKEMAHLEGLMKDLNAITTA"
     misc_feature    1493..1494
                     /label=Feature 61
     misc_feature    1543..1545
                     /gene="NEO1"
                     /gene_synonym="IGDCC2; NGN; NTN1R2"
                     /note="N-linked (GlcNAc...) asparagine.
                     /evidence=ECO:0000255; propagated from UniProtKB/Swiss-Prot
                     (Q92859.2); glycosylation site"
     primer_bind     2093..2109
                     /label=B21F-SF
     primer_bind     complement(2914..2930)
                     /label=B21B-NEO1-SR
     primer_bind     complement(3716..3756)
                     /label=B21F-BsrGI-R
     misc_feature    3736..3741
                     /label=BsrGI
     CDS             3742..3807
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3747..3835)
                     /label=296
     terminator      3811..4242
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     primer_bind     3811..3836
                     /label=#56-F-W3SL-5F1
     primer_bind     complement(3890..3913)
                     /label=R-Seq-W3SL-5R1
     misc_feature    4285..4425
                     /label=R-ITR
     rep_origin      4428..4883
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4575..4584)
                     /label=F-Seq-Cre-3F1
     promoter        5165..5269
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5270..6130
                     /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      6301..6889
                     /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 aggtcttgat
     1501 cgggggactg agtataattt ccgagtggct gctctaacaa tcaatggtac aggcccggca
     1561 actgactggc tgtctgctga aacttttgaa agtgacctag atgaaactcg tgttcctgaa
     1621 gtgcctagct ctcttcacgt acgcccgctc gttactagca tcgtagtgag ctggactcct
     1681 ccagagaatc agaacattgt ggtcagaggt tacgccattg gttatggcat tggcagccct
     1741 catgcccaga ccatcaaagt ggactataaa cagcgctatt acaccattga aaatctggat
     1801 cccagctctc actatgtgat taccctgaaa gcatttaata acgtgggtga aggcatcccc
     1861 ctgtatgaga gtgctgtgac caggcctcac acagtgccag atcccactcc catgatgcca
     1921 ccagtgggag ttcaggcttc cattctgagt catgacacca tcaggattac gtgggcagac
     1981 aactcgctgc ccaagcacca gaagattaca gactcccgat actacaccgt ccgatggaaa
     2041 accaacatcc cagcaaacac caagtacaag aatgcaaatg caaccacttt gagttatttg
     2101 gtgactggtt taaagccgaa tacactctat gaattctctg tgatggtgac caaaggtcga
     2161 agatcaagta catggagtat gacagcccat gggaccacct ttgaattagt tccgacttct
     2221 ccacccaagg atgtgactgt tgtgagtaaa gaggggaaac ctaagaccat aattgtgaat
     2281 tggcagcctc cctccgaagc caatggcaaa attacaggtt acatcatata ttacagtaca
     2341 gatgtgaatg cagagataca tgactgggtt attgagcctg ttgtgggaaa cagactgact
     2401 caccagatac aagagttaac tcttgacaca ccatactact tcaaaatcca ggcacggaac
     2461 tcaaagggca tgggacccat gtctgaagct gtccaattca gaacacctaa agcggactcc
     2521 tctgataaaa tgcctaatga tcaagcctca gggtctggag ggaaaggaag ccggctgcca
     2581 gacctaggat ccgactacaa acctccaatg agcggcagta acagccctca tgggagcccc
     2641 acctctcctc tggacagtaa tatgctgctg gtcataattg tttctgttgg cgtcatcacc
     2701 atcgtggtgg ttgtgattat cgctgtcttt tgtacccgtc gtaccacctc tcaccagaaa
     2761 aagaaacgag ctgcctgcaa atcagtgaat ggctctcata agtacaaagg gaattccaaa
     2821 gatgtgaaac ctccagatct ctggatccat catgagagac tggagctgaa acccattgat
     2881 aagtctccag acccaaaccc catcatgact gatactccaa ttcctcgcaa ctctcaagat
     2941 atcacaccag ttgacaactc catggacagc aatatccatc aaaggcgaaa ttcatacaga
     3001 gggcatgagt cagaggacag catgtctaca ctggctggaa ggcgaggaat gagaccaaaa
     3061 atgatgatgc cctttgactc ccagccaccc cagcctgtga ttagtgccca tcccatccat
     3121 tccctcgata accctcacca tcatttccac tccagcagcc tcgcttctcc agctcgcagt
     3181 catctctacc acccgggcag cccatggccc attggcacat ccatgtccct ttcagacagg
     3241 gccaattcca cagaatccgt tcgaaatacc cccagcactg acaccatgcc agcctcttcg
     3301 tctcaaacat gctgcactga tcaccaggac cctgaaggtg ctaccagctc ctcttacttg
     3361 gccagctccc aagaggaaga ttcaggccag agtcttccca ctgcccatgt tcgcccttcc
     3421 cacccattga agagcttcgc cgtgccagca atcccgcctc caggacctcc cacctatgat
     3481 cctgcattgc caagcacacc attactgtcc cagcaagctc tgaaccatca cattcactca
     3541 gtgaagacag cctccatcgg gactctagga aggagccggc ctcctatgcc agtggttgtt
     3601 cccagtgccc ctgaagtgca ggagaccaca aggatgttgg aagactccga gagtagctat
     3661 gaaccagatg agctgaccaa agagatggcc cacctggaag gactaatgaa ggacctaaac
     3721 gctatcacaa cagcatgtac agactacaaa gaccatgacg gtgattataa agatcatgac
     3781 atcgactaca aggatgacga tgacaagtaa ctcgagataa tcaacctctg gattacaaaa
     3841 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     3901 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     3961 tgtataaatc ctggttagtt cttgccacgg cggaactcat cgccgcctgc cttgcccgct
     4021 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgtttatt tgtgaaattt
     4081 gtgatgctat tgctttattt gtaaccatct agctttattt gtgaaatttg tgatgctatt
     4141 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     4201 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagatctgg agaccggtgt
     4261 ggcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     4321 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     4381 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     4441 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     4501 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4561 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4621 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4681 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     4741 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4801 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     4861 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4921 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     4981 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     5041 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     5101 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     5161 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     5221 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5281 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5341 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5401 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5461 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5521 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5581 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5641 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5701 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5761 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5821 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5881 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5941 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6001 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6061 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6121 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6181 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6241 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6301 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6361 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6421 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6481 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6541 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6601 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6661 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6721 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6781 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6841 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6901 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6961 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7021 gt
//
