LOCUS       sequence-208711-        7630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG tagged LanCL2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7630)
  AUTHORS   Zeng M, van der Donk WA, Chen J
  TITLE     Lanthionine synthetase C-like protein 2 (LanCL2) is a novel 
            regulator of Akt.
  JOURNAL   Mol Biol Cell. 2014 Dec 1;25(24):3954-61. doi: 
            10.1091/mbc.E14-01-0004. Epub 2014 Oct 1.
  PUBMED    25273559
REFERENCE   2  (bases 1 to 7630)
  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"; doi: 
            "10.1091/mbc.E14-01-0004"; journalName: "Mol Biol Cell"; date: 
            "2014-12-1- 1"; volume: "25"; issue: "24"; pages: "3954-61"
COMMENT     Sequence Label: p3×FLAG-CMV-14-LanCL2
FEATURES             Location/Qualifiers
     source          1..7630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1996..2061
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2072..2694
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2271..2290)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2718..2738)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2736..3033
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3087..3881
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3141..3160)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3751..3770
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4533..4667
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4570..4589)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4624..4643
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4672..4691)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4673..4691)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4705..4721)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4705..4721)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4718..4740)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4729..4745
                     /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)."
     promoter        complement(join(4753..4759,4760..4777,4778..4783))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4798..4819
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4936..4953)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5107..5695)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5187..5206)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5866..6657,6658..6726))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6489..6508
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6727..6831)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6858..7313
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6945..6964)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7155..7176
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7439..7461
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7453..7470
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7454..7470
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattaagc ttatgggcga gaccatgtca aagaggctga agctccacct gggaggggag
      661 gcagaaatgg aggaacgggc gttcgtcaac cccttcccgg actacgaggc cgccgccggg
      721 gcgctgctcg cctccggagc ggccgaagag acaggctgtg ttcgtccccc ggccaccccg
      781 gatgagcccg gcctcccttt tcatcaggac gggaagatca ttcataattt cataagacgg
      841 atccagacca aaattaaaga tcttctgcag caaatggaag aagggctgaa gacagctgat
      901 ccccatgact gctctgctta tactggctgg acaggcatag cccttttgta cctgcagttg
      961 taccgggtca catgtgacca aacctacctg ctccgatccc tggattacgt aaaaagaaca
     1021 cttcggaatc tgaatggccg cagggtcacc ttcctctgtg gggatgctgg ccccctggct
     1081 gttggagctg tgatttatca caaactcaga agtgactgtg agtcccagga atgtgtcaca
     1141 aaacttttgc agctccagag atcggttgtc tgccaagaat cagaccttcc tgatgagctg
     1201 ctttatggac gggcaggtta tctgtatgcc ttactgtacc tgaacacaga gataggtcca
     1261 ggcaccgtgt gtgagtcagc tattaaagag gtagtcaatg ctattattga atcgggtaag
     1321 actttgtcaa gggaagaaag aaaaacggag cgctgcccgc tgttgtacca gtggcaccgg
     1381 aagcagtacg ttggagcagc ccatggcatg gctggaattt actatatgtt aatgcagccg
     1441 gcagcaaaag tggaccaaga aaccttgaca gaaatggtga aacccagtat tgattatgtg
     1501 cgccacaaaa aattccgatc tgggaattac ccatcatcat taagcaatga aacagaccgg
     1561 ctggtgcact ggtgccacgg cgccccgggg gtcatccaca tgctcatgca ggcgtacaag
     1621 gtctttaagg aggagaagta cttgaaagag gccatggagt gtagcgatgt gatttggcag
     1681 cgaggtttgc tgcggaaggg ctacgggata tgccatggga ctgctggcaa cggctattcc
     1741 ttcctgtccc tttaccgtct cacgcaggat aagaagtacc tctaccgagc ttgcaagttt
     1801 gcagagtggt gtctagatta cggagcacac gggtgccgca ttcctgacag accctattcg
     1861 ctctttgaag gcatggctgg cgctattcac tttctctctg atgtcctggg accagagaca
     1921 tcacggtttc cagcatttga acttgactct tcgaagaggg attcggtacc agtcgactct
     1981 agaggatccc gggctgacta caaagaccat gacggtgatt ataaagatca tgacatcgac
     2041 tacaaggatg acgatgacaa gtagtgatcc cgggtggcat ccctgtgacc cctccccagt
     2101 gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt
     2161 aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt
     2221 ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga
     2281 accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca
     2341 agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct
     2401 cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc
     2461 caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc
     2521 gtgaaccact gctcccttcc ctgtccttct gattttaaaa taactatacc agcaggagga
     2581 cgtccagaca cagcataggc tacctggcca tgcccaaccg gtgggacatt tgagttgctt
     2641 gcttggcact gtcctctcat gcgttgggtc cactcagtag atgcctgttg aattgggtac
     2701 gcggccagct tggctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2761 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     2821 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     2881 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     2941 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3001 gctattccag aagtagtgag gaggcttttt tggaggaatt gatcagcttg ggatctgatc
     3061 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3121 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3181 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3241 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3301 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3361 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3421 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3481 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3541 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3601 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3661 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3721 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3781 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3841 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     3901 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     3961 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4021 cggggatctc atgctggagt tcttcgccca ccccgggctc gatcccctcg cgagttggtt
     4081 cagctgctgc ctgaggctgg acgacctcgc ggagttctac cggcagtgca aatccgtcgg
     4141 catccaggaa accagcagcg gctatccgcg catccatgcc cccgaactgc aggagtgggg
     4201 aggcacgatg gccgctttgg tcgacccgga cgggacgctc ctgcgcctga tacagaacga
     4261 attgcttgca ggcatctcat gagtgtgtct tcccgttttc cgcctgaggt cactgcgtgg
     4321 atggagcgct ggcgcctgct gcgcgacggc gagctgctca ccacccactc gccaagctgg
     4381 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgccgatc ataatcagcc
     4441 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     4501 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     4561 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4621 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaat tccctatagt
     4681 gagtcgtatt aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct
     4741 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4801 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4861 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4921 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4981 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5041 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5101 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5161 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5221 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5281 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5341 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5401 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5461 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5521 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5581 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5641 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5701 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5761 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5821 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5881 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5941 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6001 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6061 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6121 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6181 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6241 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6301 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6361 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6421 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6481 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6541 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6601 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6661 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6721 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6781 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6841 ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     6901 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     6961 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     7021 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     7081 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     7141 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     7201 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     7261 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttccattcg
     7321 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     7381 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     7441 cagtcacgac gttgtaaaac gacggccagt gccaagctga tctatacatt gaatcaatat
     7501 tggcaattag ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca
     7561 ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga
     7621 ccgccatgtt
//
