LOCUS       pSL48_CFAP36(pig        7520 bp    DNA     circular SYN 23-JUN-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7520)
  AUTHORS   MRC Lab University of Dundee Allison Bridges
  TITLE     Direct Submission
  JOURNAL   Exported Apr 9, 2025 from SnapGene 8.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          5317..6342
                     /chromosome="3"
                     /mol_type="mRNA"
                     /db_xref="taxon:9823"
                     /organism="Sus scrofa"
     promoter        104..208
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(209..277,278..1069)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1240..1828
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2014..2156
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2258..2449)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3761..4843)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4844..4921)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5230..5249
                     /label=T7
     promoter        5230..5248
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5249..5273
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(5272..5319)
                     /label=oSL27_p_rev
     RBS             5288..5310
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5317..6342
                     /codon_start=1
                     /gene="CFAP36"
                     /gene_synonym="CCDC104"
                     /product="cilia- and flagella-associated protein 36"
                     /label=CFAP36
                     /note="coiled-coil domain containing 104"
                     /db_xref="GeneID:100513146"
                     /db_xref="VGNC:VGNC:86599"
                     /protein_id="NP_001231704.1"
                     /translation="MAAEEDEVEWVVESIAGFLRGPDWSIPILDFVEQKCEVFDDEEES
                     KLTYTEIHQEYKELVEKLLESYLKEIGINEDQFQEACTSPLAKTHTSQAILQPVLAAED
                     FTIFKAMMVQKNIEMQLQAIRIIQERNGVLPDCLTDGSDVVSDLEQEEMKILREVLRKE
                     KEEYDQEEERKRKNQLSEAKTEEPSMHANETVKMNNSQGDGEHFAHPPSEVKVHFANQS
                     VQPLTTKLEMLPETSTLPKKGLKIPGFERASIEGPIANLSTLGTEELRQREHYLKQKRD
                     KLMSMRKDMRTKQIQNLEEKGTPVGELEEMTEKPEMTAEEKQTLLKRRLLAEKLKEEVI
                     NK"
     misc_feature    5317..5718
                     /label=BART-like domain
     primer_bind     5324..5357
                     /label=oSL45_fwd
     primer_bind     complement(5324..5357)
                     /label=oSL45_rev
     primer_bind     5569..5613
                     /label=oSL46_fwd
     primer_bind     complement(5569..5613)
                     /label=oSL46_rev
     primer_bind     5659..5710
                     /label=oSL47_fwd
     primer_bind     complement(5659..5710)
                     /label=oSL47_rev
     misc_feature    5761..5865
                     /label=UIMs (deg)
     primer_bind     complement(5774..5827)
                     /label=oSL48_rev
     primer_bind     5774..5827
                     /label=oSL48_fwd
     primer_bind     complement(5825..5877)
                     /label=oSL49_rev
     primer_bind     5825..5877
                     /label=oSL49_fwd
     primer_bind     complement(5834..5874)
                     /label=oSL43_rev
     primer_bind     complement(5834..5874)
                     /label=oSL44_rev
     misc_feature    6112..6201
                     /label=Conserved helix
     misc_feature    6271..6342
                     /label=Conserved helix
     primer_bind     complement(6341..6364)
                     /label=oSL43_rev
     primer_bind     6343..6370
                     /label=oSL27_GFP_fwd
     CDS             6343..6354
                     /codon_start=3
                     /locus_tag="GSQ linker"
                     /label=Linker
                     /label=GSQ linker
                     /translation="GSGS"
     CDS             6355..6378
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=PreScission site
                     /translation="LEVLFQGP"
     CDS             6379..6417
                     /codon_start=3
                     /locus_tag="GSQ linker"
                     /label=Linker
                     /label=GSQ linker
                     /translation="GGRGGSGGQGSGS"
     CDS             6418..7131
                     /codon_start=1
                     /product="A Novel Ultra-Stable, Monomeric Green Fluorescent
                     Protein For Direct Volumetric Imaging of Whole Organs Using
                     CLARITY
                     Scott Dj, Gunn Nj, Yong Kj, Wimmer Vc, Veldhuis Na, Challis
                     Lm, Haidar M, Petrou S, Bathgate Rad, Griffin Mdw
                     (2018). Scientific Reports, 8(1) , 667. doi: 
                     10.1038/s41598-017-18045-y"
                     /label=muGFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVLCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     YFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     DVTAAGITHGMDELYK"
     primer_bind     6418..6451
                     /label=oSL44_fwd
     primer_bind     complement(6448..6476)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     complement(7099..7167)
                     /label=oSL27_GFP_rev
     CDS             7132..7179
                     /codon_start=1
                     /label=10His
                     /translation="GGSKHHHHHHHHHHGE"
     primer_bind     7144..7196
                     /label=oSL27_p_fwd
     primer_bind     7184..7202
                     /label=oSL2_p_fwd
     primer_bind     complement(7247..7265)
                     /label=T7 term
     terminator      7261..7308
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(7483..7488,7489..7505,7506..7511))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
       61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      181 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
      481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
      541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
      721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
      781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
      841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
      901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
      961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     1981 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2041 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc
     2101 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     2161 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2221 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2281 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     2341 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     2401 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     2461 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     2521 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     2581 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     2641 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     2701 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     2761 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     2821 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     2881 tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga
     2941 tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg
     3001 tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc
     3061 cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg
     3121 acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt
     3181 ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccag tgatcgaagt
     3241 taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg
     3301 cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat
     3361 aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc
     3421 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt
     3481 gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat
     3541 cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg
     3601 tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg
     3661 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg
     3721 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3781 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3841 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
     3901 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     3961 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
     4021 cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc
     4081 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     4141 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     4201 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     4261 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     4321 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     4381 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     4441 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     4501 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     4561 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     4621 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     4681 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     4741 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     4801 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     4861 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt
     4921 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     4981 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     5041 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     5101 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     5161 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
     5221 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
     5281 aaataatttt gtttaacttt aagaaggaga tataccatgg ctgcagaaga ggacgaggtg
     5341 gaatgggtgg tggagagcat tgcgggtttc ctgcgaggcc cggactggtc catccccatc
     5401 ttggactttg tggagcagaa atgtgaagtt tttgatgatg aagaagagag caaattgacc
     5461 tatacagaga ttcatcagga atacaaagaa ctagttgaaa agctgttaga aagttacctc
     5521 aaagaaattg gaattaatga agatcagttt caagaagcat gcacttctcc tcttgcaaag
     5581 acccatacct cacaggccat tttgcaacct gtgttggcag cagaagattt tactatcttt
     5641 aaagcgatga tggtccagaa aaacattgaa atgcagctgc aagccattcg gataattcaa
     5701 gaaagaaatg gtgtattacc tgactgctta acagatggtt ctgatgtggt cagtgacctt
     5761 gaacaggaag agatgaaaat cctgagggaa gttcttagaa aagagaaaga ggaatatgac
     5821 caggaggaag aaaggaagag aaaaaatcag ttatcagagg ctaaaacaga agagccctcg
     5881 atgcatgcca atgaaactgt aaaaatgaat aattcccaag gggatggtga acactttgca
     5941 cacccaccct cagaagttaa agtgcatttt gctaatcagt cagtacaacc tttgacaaca
     6001 aagctggaaa tgttgcctga aacttctacc ctcccgaaaa aaggcctgaa gattcctggc
     6061 ttcgaacgtg caagcattga aggaccaata gcaaatttat caacacttgg aacagaagag
     6121 ctccggcaac gagaacatta tctaaagcag aagagagata aattgatgtc catgagaaag
     6181 gatatgagga ctaagcagat acaaaatttg gaggagaaag gaacacctgt tggcgagtta
     6241 gaggaaatga cagagaaacc agaaatgaca gcagaggaga agcaaacatt actaaagagg
     6301 agattgcttg cagagaaact taaagaagaa gttattaata agggatctgg ctctctggaa
     6361 gttctgttcc aggggcccgg cggccgcggg ggatctggcg ggcaaggatc tggctctatg
     6421 agcaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat
     6481 gttaatgggc acaaattttc tgtccgtgga gagggtgaag gtgatgctac aaacggaaaa
     6541 ctcaccctta aatttatttg cactactgga aaactacctg ttccgtggcc aacacttgtc
     6601 actactctga cctatggtgt tctatgcttt tcccgttatc cggatcacat gaaacggcat
     6661 gactttttca agagtgccat gcccgaaggt tatgtacagg aacgcactat atctttcaaa
     6721 gatgacggga cctacaagac gcgtgctgaa gtcaagtttg aaggtgatac ccttgttaat
     6781 cgtatcgagt taaaaggtat tgattttaaa gaagatggaa acattctcgg acacaaactg
     6841 gagtacaact ttaactcaca caatgtatac atcacggcag acaaacaaaa gaatggaatc
     6901 aaagcgtact tcaaaattcg ccacaacgtt gaagatggtt ccgttcaact agcagaccat
     6961 tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctc
     7021 tcgacacaat ctgttctttc gaaagatccc aacgaaaagc gtgaccacat ggtccttctt
     7081 gaggatgtaa ctgctgctgg gattacacat ggcatggatg aactctacaa aggaggatct
     7141 aagcatcacc atcatcatca ccatcaccac catggagagt gagcggccgc agatccagat
     7201 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     7261 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     7321 actatatccg gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta
     7381 cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg
     7441 gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga
     7501 taagctgtca aacatgagaa
//
