LOCUS       Exported                6621 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 6621)
  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..6621
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1493..3322
                     /organism="Homo sapiens"
                     /chromosome="1"
                     /map="1q22"
                     /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..1508
                     /label=B35F-NheI-F
     primer_bind     1477..1492
                     /label=Primer 10
     misc_feature    1492
                     /label=NheI?
     CDS             1493..3322
                     /codon_start=1
                     /gene="IQGAP3"
                     /product="ras GTPase-activating-like protein IQGAP3"
                     /label=IQGAP3
                     /db_xref="GI:1519313212"
                     /db_xref="CCDS:CCDS1144.1"
                     /db_xref="GeneID:128239"
                     /db_xref="HGNC:HGNC:20669"
                     /protein_id="NP_839943.3"
                     /translation="VEQPQDVVTGNPTVVRLVVRFYRNGRGQSALQEILGKVIQDVLED
                     KVLSVHTDPVHLYKNWINQTEAQTGQRSHLPYDVTPEQALSHPEVQRRLDIALRNLLAM
                     TDKFLLAITSSVDQIPYGMRYVAKVLKATLAEKFPDATDSEVYKVVGNLLYYRFLNPAV
                     VAPDAFDIVAMAAGGALAAPQRHALGAVAQLLQHAAAGKAFSGQSQHLRVLNDYLEETH
                     LKFRKFIHRACQVPEPEERFAVDEYSDMVAVAKPMVYITVGELVNTHRLLLEHQDCIAP
                     DHQDPLHELLEDLGELPTIPDLIGESIAADGHTDLSKLEVSLTLTNKFEGLEADADDSN
                     TRSLLLSTKQLLADIIQFHPGDTLKEILSLSASREQEAAHKQLMSRRQACTAQTPEPLR
                     RHRSLTAHSLLPLAEKQRRVLRNLRRLEALGLVSARNGYQGLVDELAKDIRNQHRHRHR
                     RKAELVKLQATLQGLSTKTTFYEEQGDYYSQYIRACLDHLAPDSKSSGKGKKQPSLHYT
                     AAQLLEKGVLVEIEDLPASHFRNVIFDITPGDEAGKFEVNAKFLGVDMERFQLHYQDLL
                     QLQYEGVAVMKLFNKAKVNVNLLIFLLNKKFLRK"
     misc_feature    1493..1494
                     /label=Feature 49
     primer_bind     1628..1644
                     /label=B35B-IQGAP3-SF
     primer_bind     2067..2083
                     /label=B35F-SF
     primer_bind     2250..2267
                     /label=B35F-BsrGI-1F
     primer_bind     complement(3302..3343)
                     /label=B35F-BsrGI-R
     primer_bind     complement(3309..3346)
                     /label=B35F-BsrGI-1R
     primer_bind     complement(3309..3328)
                     /label=Primer 1
     misc_feature    3323..3328
                     /label=BsrGI
     CDS             3329..3394
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3334..3434)
                     /label=296
     terminator      3410..3841
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     primer_bind     3410..3435
                     /label=#56-F-W3SL-5F1
     primer_bind     complement(3489..3512)
                     /label=R-Seq-W3SL-5R1
     misc_feature    3884..4024
                     /label=R-ITR
     rep_origin      4027..4482
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4174..4183)
                     /label=F-Seq-Cre-3F1
     promoter        4764..4868
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4869..5729
                     /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      5900..6488
                     /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 aggtggagca
     1501 gccccaggac gtggtgacag gcaacccaac agtggtgagg ctggtggtga gattctaccg
     1561 taatgggcgg ggacagagtg ccctgcagga gattctgggc aaggttatcc aggatgtgct
     1621 agaagacaaa gtgctcagcg tccacacaga ccctgtccac ctctataaga actggatcaa
     1681 ccagactgag gcccagacag ggcagcgcag ccatctccca tatgatgtca ccccggagca
     1741 ggccttgagc caccccgagg tccagagacg actggacatc gccctacgca acctcctcgc
     1801 catgactgat aagttccttt tagccatcac ctcatctgtg gaccaaattc cgtatgggat
     1861 gcgatatgtg gccaaagtcc tgaaggcaac tctggcagag aaattccctg acgccacaga
     1921 cagcgaggtc tataaggtgg tcgggaacct cctgtactac cgcttcctga acccagctgt
     1981 ggtggctcct gacgccttcg acattgtggc catggcagct ggtggagccc tggctgcccc
     2041 ccagcgccat gccctggggg ctgtggctca gctcctacag cacgctgcgg ctggcaaggc
     2101 cttctctggg cagagccagc acctacgggt cctgaatgac tatctggagg aaacacacct
     2161 caagttcagg aagttcatcc atagagcctg ccaggtgcca gagccagagg agcgttttgc
     2221 agtggacgag tactcagaca tggtggctgt ggccaaaccc atggtgtaca tcaccgtggg
     2281 ggagctggtc aacacgcaca ggctgttgct ggagcaccag gactgcattg cccctgatca
     2341 ccaagacccc ctgcatgagc tcctggagga tcttggggag ctgcccacca tccctgacct
     2401 tattggtgag agcatcgctg cagatgggca cacggacctg agcaagctag aagtgtccct
     2461 gacgctgacc aacaagtttg aaggactaga ggcagatgct gatgactcca acacccgtag
     2521 cctgcttctg agcaccaagc agctgttggc cgatatcata cagttccatc ctggggacac
     2581 cctcaaggag atcctgtccc tctcggcttc cagagagcaa gaagcagccc acaagcagct
     2641 gatgagccga cgccaggcct gtacagccca gacaccggag ccactgcgac gacaccgctc
     2701 actgacagct cactccctcc tgccactggc agagaagcag cggcgcgtcc tgcggaacct
     2761 acgccgactt gaagccctgg ggttggtcag cgccagaaat ggctaccagg ggctagtgga
     2821 cgagctggcc aaggacatcc gcaaccagca cagacacagg cacaggcgga aggcagagct
     2881 ggtgaagctg caggccacat tacagggcct gagcactaag accaccttct atgaggagca
     2941 gggtgactac tacagccagt acatccgggc ctgcctggac cacctggccc ccgactccaa
     3001 gagttctggg aaggggaaga agcagccttc tcttcattac actgctgctc agctcctgga
     3061 aaagggtgtc ttggtggaaa ttgaagatct tcccgcctct cacttcagaa acgtcatctt
     3121 tgacatcacg ccgggagatg aggcaggaaa gtttgaagta aatgccaagt tcctgggtgt
     3181 ggacatggag cgatttcagc ttcactatca ggatctcctg cagctccagt atgagggtgt
     3241 ggctgtcatg aaactcttca acaaggccaa agtcaatgtc aaccttctca tcttcctcct
     3301 caacaagaag tttttgcgga agtgtacaga ctacaaagac catgacggtg attataaaga
     3361 tcatgacatc gactacaagg atgacgatga caagtaagaa ttctgatgac tcgagataat
     3421 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     3481 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     3541 gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc ggaactcatc
     3601 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3661 gtgtttattt gtgaaatttg tgatgctatt gctttatttg taaccatcta gctttatttg
     3721 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3781 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta
     3841 aagatctgga gaccggtgtg gcacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     3901 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     3961 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     4021 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4081 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4141 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     4201 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4261 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     4321 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4381 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     4441 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     4501 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     4561 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     4621 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4681 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4741 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     4801 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4861 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4921 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4981 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5041 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5101 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5161 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5221 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5281 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5341 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5401 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5461 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5521 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5581 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5641 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5701 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5761 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5821 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5881 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5941 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6001 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6061 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6121 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6181 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6241 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6301 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6361 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6421 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6481 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6541 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6601 gctggccttt tgctcacatg t
//
