LOCUS       Exported                6429 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 6429)
  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..6429
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1493..3142
                     /organism="Homo sapiens"
                     /chromosome="5"
                     /map="5q12.3"
                     /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=B14F-NheI-F
     primer_bind     1477..1492
                     /label=Primer 10
     misc_feature    1492
                     /label=NheI?
     CDS             1493..3142
                     /codon_start=1
                     /gene="ERBIN"
                     /gene_synonym="ERBB2IP; HEL-S-78; LAP2"
                     /product="erbin isoform 1"
                     /label=ERBIN
                     /note="isoform 1 is encoded by transcript variant 1; 
                     densin-180-like protein; protein LAP2; epididymis secretory
                     protein Li 78"
                     /db_xref="GI:358679311"
                     /db_xref="CCDS:CCDS58953.1"
                     /db_xref="GeneID:55914"
                     /db_xref="HGNC:HGNC:15842"
                     /db_xref="MIM:606944"
                     /protein_id="NP_001240626.1"
                     /translation="VNGHSEETSQSPNRTEPHDSDCSVDLGISKSTEDLSPQKSGPVGS
                     VVKSHSITNMEIGGLKIYDILSDNGPQQPSTTVKITSAVDGKNIVRSKSATLLYDQPLQ
                     VFTGSSSSSDLISGTKAIFKFDSNHNPEEPNIIRGPTSGPQSAPQIYGPPQYNIQYSSS
                     AAVKDTLWHSKQNPQIDHASFPPQLLPRSESTENQSYAKHSANMNFSNHNNVRANTAYH
                     LHQRLGPARHGEMWAISPNDRLIPAVTRSTIQRQSSVSSTASVNLGDPGSTRRAQIPEG
                     DYLSYREFHSAGRTPPMMPGSQRPLSARTYSIDGPNASRPQSARPSINEIPERTMSVSD
                     FNYSRTSPSKRPNARVGSEHSLLDPPGKSKVPRDWREQVLRHIEAKKLEKMPLSNGQMG
                     QPLRPQANYSQIHHPPQASVARHPSREQLIDYLMLKVAHQPPYTQPHCSPRQGHELAKQ
                     EIRVRVEKDPELGFSISGGVGGRGNPFRPDDDGIFVTRVQPEGPASKLLQPGDKIIQAN
                     GYSFINIEHGQAVSLLKTFQNTVELIIVREVSS"
     misc_feature    1493..1494
                     /label=Feature 80
     primer_bind     1899..1915
                     /label=B14B-ERBIN-SF
     primer_bind     2064..2080
                     /label=B14F-SF
     primer_bind     complement(3122..3163)
                     /label=B14F-BsrGI-R
     misc_feature    3143..3148
                     /label=BsrGI
     CDS             3149..3214
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3154..3242)
                     /label=296
     terminator      3218..3649
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     primer_bind     3218..3243
                     /label=#56-F-W3SL-5F1
     primer_bind     complement(3297..3320)
                     /label=R-Seq-W3SL-5R1
     misc_feature    3692..3832
                     /label=R-ITR
     rep_origin      3835..4290
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3982..3991)
                     /label=F-Seq-Cre-3F1
     promoter        4572..4676
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4677..5537
                     /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      5708..6296
                     /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 aggtaaatgg
     1501 acattctgag gaaacttccc agtctcctaa taggactgaa ccacatgaca gtgattgttc
     1561 tgttgactta ggtatttcca aaagcactga agatctctcc cctcagaaaa gtggtccagt
     1621 tggatctgtt gtgaaatctc atagcataac taatatggag attggagggc taaaaatcta
     1681 tgatattctt agtgataatg gacctcagca gccaagtaca accgttaaaa tcacatctgc
     1741 tgttgatgga aaaaatatag tcaggagcaa gtctgccaca ctgttgtatg atcaaccatt
     1801 gcaggtattt actggttctt cctcatcttc tgatttaata tcaggaacaa aggcaatttt
     1861 caagtttgat tcaaatcata atcccgaaga gccaaatata ataagaggcc ccacaagtgg
     1921 cccacaatct gcacctcaaa tatatggtcc tccacagtat aatatccaat acagtagcag
     1981 tgctgcagtc aaagacactt tgtggcactc caaacaaaat ccccaaatag accatgccag
     2041 ttttcctcct cagctccttc ctagatcaga gagcacagaa aatcaaagtt atgctaaaca
     2101 ttctgccaat atgaatttct ctaatcataa caatgttcga gctaatactg cataccattt
     2161 acatcagaga cttggcccag caagacatgg ggaaatgtgg gccatctcac caaacgaccg
     2221 acttattcct gcagtaactc gaagtacaat ccagcgacaa agtagtgtgt cctccacagc
     2281 ctctgtaaat cttggtgatc caggctctac aaggcgggct cagattcctg aaggagatta
     2341 tttatcatac agagagttcc actcagcggg aagaactcct ccaatgatgc caggatcaca
     2401 gagacccctt tctgcacgaa catacagcat agatggtcca aatgcatcaa gacctcagag
     2461 tgctcgaccc tctattaatg aaataccaga gagaactatg tcagttagtg atttcaatta
     2521 ttcacggact agtccttcaa aaagaccaaa tgcaagggtt ggttctgagc attctttatt
     2581 agatcctcca ggaaaaagta aagttcctcg tgactggaga gaacaagtac ttcgacatat
     2641 tgaagccaaa aagttagaaa agatgccttt gagtaatgga cagatgggcc agcctctcag
     2701 gcctcaggca aattatagtc aaatacatca cccccctcag gcatctgtgg caaggcatcc
     2761 ctctagagaa caactaattg attacttgat gctgaaagtg gcccaccagc ctccatatac
     2821 acagccccat tgttctccta gacaaggcca tgaactggca aaacaagaga ttcgagtgag
     2881 ggttgaaaag gatccagaac ttggatttag catatcaggt ggtgtcgggg gtagaggaaa
     2941 cccattcaga cctgatgatg atggtatatt tgtaacaagg gtacaacctg aaggaccagc
     3001 atcaaaatta ctgcagccag gtgataaaat tattcaggct aatggctaca gttttataaa
     3061 tattgaacat ggacaagcag tgtccttgct aaaaactttc cagaatacag ttgaactcat
     3121 cattgtacga gaagtttcct catgtacaga ctacaaagac catgacggtg attataaaga
     3181 tcatgacatc gactacaagg atgacgatga caagtaactc gagataatca acctctggat
     3241 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3301 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3361 tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc cgcctgcctt
     3421 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gtttatttgt
     3481 gaaatttgtg atgctattgc tttatttgta accatctagc tttatttgtg aaatttgtga
     3541 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3601 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gatctggaga
     3661 ccggtgtggc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     3721 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     3781 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     3841 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     3901 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3961 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     4021 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4081 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     4141 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4201 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg
     4261 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     4321 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     4381 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     4441 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4501 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     4561 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     4621 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4681 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4741 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4801 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4861 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4921 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4981 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5041 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5101 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5161 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5221 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5281 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5341 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5401 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5461 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5521 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5581 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5641 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5701 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5761 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5821 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5881 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5941 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6001 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6061 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6121 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6181 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6241 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6301 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6361 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6421 ctcacatgt
//
