LOCUS       sequence-63155-e        7793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lacritin-intein fusion plasmid coding for intact mature lacritin 
            without signal peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7793)
  AUTHORS   Wang J, Wang N, Xie J, Walton SC, McKown RL, Raab RW, Ma P, Beck SL,
            Coffman GL, Hussaini IM, Laurie GW
  TITLE     Restricted epithelial proliferation by lacritin via 
            PKCalpha-dependent NFAT and mTOR pathways.
  JOURNAL   J Cell Biol. 2006 Aug 28;174(5):689-700. Epub 2006 Aug 21.
  PUBMED    16923831
REFERENCE   2  (bases 1 to 7793)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Biol."; date: "2006-08-28"; volume: "174(5)"; pages: "689-700. Epub 
            2006 Aug 21"
COMMENT     Sequence Label: pLAC
FEATURES             Location/Qualifiers
     source          1..7793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..139
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(140..208,209..1000)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(358..377)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(1042..1555)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1237..1258)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1449..1468
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      1666..2254
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2155..2174
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2408..2425
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2447..2466)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2607..2629
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3688..4770)
                     /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"
     primer_bind     4732..4751
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4771..4848)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      5001..5087
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5184..5270
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5367..5453
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5550..5636
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             6092..6910
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae VMA1"
                     /product="5' region of the intein from the yeast Vma1 
                     subunit of the vacuolar ATPase (Chong et al., 1998)"
                     /label=Sce VMA intein 5' region
                     /translation="CFAKGTNVLMADGSIECIENIEVGNKVMGKDGRPREVIKLPRGRE
                     TMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRTPRSVRRLSRTIKGVEYFEV
                     ITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELVESYRKASNKAYFEWTIEARD
                     LSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLTIEGPKVLAYLLGLWIGDGLSD
                     RATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQVAKTVNLYSKVVRG"
     CDS             7366..7377
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             7487..7642
                     /codon_start=1
                     /gene="Bacillus circulans chiA"
                     /product="chitin binding domain from chitinase A1 (Watanabe
                     et al., 1994)"
                     /label=CBD
                     /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV
                     PALWQLQ"
     primer_bind     complement(7707..7725)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7721..7768
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 aactacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
       61 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
      121 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
      181 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
      241 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
      301 ccttgagagt tttcgccccg aagaacgttc tccaatgatg agcactttta aagttctgct
      361 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
      421 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
      481 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
      541 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
      601 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
      661 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
      721 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
      781 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
      841 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
      901 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
      961 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1021 atatatactt tagattgatt taccccggtt gataatcaga aaagccccaa aaacaggaag
     1081 attgtataag caaatattta aattgtaaac gttaatattt tgttaaaatt cgcgttaaat
     1141 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     1201 tcaaaagaat agcccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     1261 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     1321 ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     1381 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     1441 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     1501 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtaaaag
     1561 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     1621 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1681 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1741 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1801 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     1861 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     1921 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     1981 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2041 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2101 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2161 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2221 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2281 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2341 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2401 cgagcgcagc gagtcagtga gcgaggaagc tatggtgcac tctcagtaca atctgctctg
     2461 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     2521 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     2581 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     2641 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gcagcgattc
     2701 acagatgtct gcctgttcat ccgatggaag atgcctcctc tgactcgacg ggtgctgatc
     2761 ctgcccagga agctgggacc tctaagccta atgaagagat ctcaggtcca gcagaaccag
     2821 cttcaccccc agagacaacc acaacagccc aggagacttc ggcggcagca gttcagggga
     2881 cagccaaggt cacctcaagc aggcaggaac taaaccccct gaaatccata gtggagaaaa
     2941 gtatcttact aacagaacaa gcccttgcaa aagcaggaaa aggaatgcac ggaggcgtgc
     3001 caggtggaaa acaatccatc gaaaatggaa gtgaatttgc acaaaaatta ctgaagaaat
     3061 tcagtctatt aaaaccatgg gcatgagata aagcgggcca tgttaagggc ggttttttcc
     3121 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     3181 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     3241 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     3301 cagggtcaat gccagccgaa cgccagcaag acgtagccca gcgcgtcggc cgccatgccg
     3361 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga
     3421 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag
     3481 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc
     3541 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
     3601 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     3661 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3721 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3781 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     3841 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     3901 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     3961 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     4021 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     4081 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     4141 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     4201 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     4261 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     4321 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     4381 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     4441 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     4501 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     4561 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     4621 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     4681 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     4741 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     4801 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     4861 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     4921 gcaccgccgc cgcaaggaat ggtgcatgcc ggcatgccgc cctttcgtct tcaagaatta
     4981 attcccaatt ccccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     5041 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg
     5101 atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg
     5161 ccaggaatta attccccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     5221 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag
     5281 cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa
     5341 ctgccaggaa ttaattcccc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     5401 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
     5461 gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
     5521 aaactgccag gaattaattc cccaggcatc aaataaaacg aaaggctcag tcgaaagact
     5581 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     5641 cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc
     5701 cataaactgc caggaattaa ttcatggaag atgcctcctc tgactcgacg ggtgctgatc
     5761 ctgcccagga agctgggacc tctaagccta atgaagagat ctcaggtcca gcagaaccag
     5821 cttcaccccc agagacaacc acaacagccc aggagacttc ggcggcagca gttcagggga
     5881 cagccaaggt cacctcaagc aggcaggaac taaaccccct gaaatccata gtggagaaaa
     5941 gtatcttact aacagaacaa gcccttgcaa aagcaggaaa aggaatgcac ggaggcgtgc
     6001 caggtggaaa acaatccatc gaaaatggaa gtgaatttgc acaaaaatta ctgaagaaat
     6061 tcagtctatt aaaaccatgg gcatgacttc ctgctttgcc aagggtacca atgttttaat
     6121 ggcggatggg tctattgaat gtattgaaaa cattgaggtt ggtaataagg tcatgggtaa
     6181 agatggcaga cctcgtgagg taattaaatt gcccagagga agagaaacta tgtacagcgt
     6241 cgtgcagaaa agtcagcaca gagcccacaa aagtgactca agtcgtgaag tgccagaatt
     6301 actcaagttt acgtgtaatg cgacccatga gttggttgtt agaacacctc gtagtgtccg
     6361 ccgtttgtct cgtaccatta agggtgtcga atattttgaa gttattactt ttgagatggg
     6421 ccaaaagaaa gcccccgacg gtagaattgt tgagcttgtc aaggaagttt caaagagcta
     6481 cccaatatct gaggggcctg agagagccaa cgaattagta gaatcctata gaaaggcttc
     6541 aaataaagct tattttgagt ggactattga ggccagagat ctttctctgt tgggttccca
     6601 tgttcgtaaa gctacctacc agacttacgc tccaattctt tatgagaatg accacttttt
     6661 cgactacatg caaaaaagta agtttcatct caccattgaa ggtccaaaag tacttgctta
     6721 tttacttggt ttatggattg gtgatggatt gtctgacagg gcaacttttt cggttgattc
     6781 cagagatact tctttgatgg aacgtgttac tgaatatgct gaaaagttga atttgtgcgc
     6841 cgagtataag gacagaaaag aaccacaagt tgccaaaact gttaatttgt actctaaagt
     6901 tgtcagaggt aatggtattc gcaataatct taatactgag aatccattat gggacgctat
     6961 tgttggctta ggattcttga aggacggtgt caaaaatatt ccttctttct tgtctacgga
     7021 caatatcggt actcgtgaaa catttcttgc tggtctaatt gattctgatg gctatgttac
     7081 tgatgagcat ggtattaaag caacaataaa gacaattcat acttctgtca gagatggttt
     7141 ggtttccctt gctcgttctt taggcttagt agtctcggtt aacgcagaac ctgctaaggt
     7201 tgacatgaat gtcaccaaac ataaaattag ttatgctatt tatatgtctg gtggagatgt
     7261 tttgcttaac gttctttcga agtgtgccgg ctctaaaaaa ttcaggcctg ctcccgccgc
     7321 tgcttttgca cgtgagtgcc gcggatttta tttcgagtta caagaattga aggaagacga
     7381 ttattatggg attactttat ctgatgattc tgatcatcag tttttgcttg gatcccaggt
     7441 tgtcgtccat gcatgcggtg gcctgaccgg tctgaactca ggcctcacga caaatcctgg
     7501 tgtatccgct tggcaggtca acacagctta tactgcggga caattggtca catataacgg
     7561 caagacgtat aaatgtttgc agccccacac ctccttggca ggatgggaac catccaacgt
     7621 tcctgccttg tggcagcttc aatgactgca ggaaggggat ccggctgcta acaaagcccg
     7681 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     7741 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gat
//
