LOCUS       sequence-210701-        9790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP reporter of lag-2 driven by 0.5 kb promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9790)
  AUTHORS   Pekar O, Ow MC, Hui KY, Noyes MB, Hall SE, Hubbard EJA
  TITLE     Linking the environment, DAF-7/TGFbeta signaling and LAG-2/DSL 
            ligand expression in the germline stem cell niche.
  JOURNAL   Development. 2017 Aug 15;144(16):2896-2906. doi: 10.1242/dev.147660.
  PUBMED    28811311
REFERENCE   2  (bases 1 to 9790)
  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: "10.1242/dev"; 
            journalName: "Development"; date: "2017-08-15- 15"; volume: "144"; 
            issue: "16"; pages: "2896-2906"
COMMENT     Sequence Label: pGC643
FEATURES             Location/Qualifiers
     source          1..9790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1490..1501)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4654..4682)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5430..5450
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(5931..5948)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5931..5947)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7774..7796
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7834..7852)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7920..8024
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8025..8093,8094..8885)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8243..8262)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9056..9644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9545..9564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccctgtcg gattatgggc tcttctttta tcctctactt tctttctaga aaatgcaaat
       61 gttgagtcag aaattgctga atggagtaca actgcaatgg gtttagctta tgtttctgga
      121 acaattactg gtggaattgt gattgaaaag taaggttata tttcatttca ataaatgtta
      181 ccacagtata acaaaaacaa gttacattgt tcagagtatg tctgaatgta ttattattat
      241 tgtatttgtg catcagtact gtagttgtac tgaagaaggg tttaaaataa tctcaacgtt
      301 tttaagacga gttagtgaga atacggtcgc agttaggatg tttggacata gttttattag
      361 ataatcgtga actaactcac tataaaaacc cgctaactca ttgaataaga tagttttaaa
      421 cgttcaaaat ttgaagacaa aattgtatga ataagtttac agagttggtc gacgcaagct
      481 tctccttctc ttcacatttc tcaataattt agctctaatt gcattcgtat tcttcgcaaa
      541 aatacgtatt ctgattgacc cgatgaaata cggatgccta gttgcattga tcatctacgg
      601 ttatacttat gggtgaggtt tttcaaatat attttgattt aatcgaataa tgaaatttca
      661 gaactggagt tggtccaatt tcgtggttta tatcctctga actggtacct caaaaacatc
      721 gtagtattgc tcaatctgtg gcatattcaa tgaatactgt aatggtggtt atcagtacat
      781 ttacagtatt acctctatat tccgttattg gtagttatgc tttccttatt ctatattcaa
      841 ttccttcatt tataagtatg cttatattat ttcgctactt accggaaacc aaaggacgag
      901 agattcatga tattgtgaat gaacttaaaa aaaaatgaaa acctttgaaa acgttcatac
      961 ggccgaaatt ttacttgatc gagccaattg aacttttgtt aagacttaca gaaaagtgag
     1021 agattttgtt acaaaaaagt gagaaaaaag cgaaaatgaa caggaaaaag aagaaaaata
     1081 cacccgaagt atcatataaa aattaaaaag attgtttgac ctggcggaac ttttttgcaa
     1141 aaactctgaa tatccatggc acatgcttcg tgcaaaacag gatccaaatg aatatcaaca
     1201 agagattctt gaagacgacg agccacttgc tcagagcatt gcttatctcg aatgagaccc
     1261 ttgttaaaat catttttcaa acattatatg acattgtcga aatgtcctcc tgattccatg
     1321 atggtagcaa actcacttcg tctttgagcc aatttatcca agtccttgta aaaagtattc
     1381 gaaaattgtt aacggataaa atgtttatta taatatcaaa aacaatttgt cagttgacca
     1441 ctttttgata tatttgacag aaaccgggat gaattggctc aaagttaggg cgcccttcta
     1501 ttacagggtt tctgataaca aaccggttat taactcccaa caagggatga tttcaattca
     1561 tcatgctcaa attgacccaa attaagttac atgacaaatt catcgccctt ttcaactctt
     1621 cttggctcat catctgttat tctgttccta ttctgtctgc accctatacc cctttgcata
     1681 ctctctcgtc attcctttct tggatagtgt cttcttctcc ccagctctgc tacttctatg
     1741 acttgcgccc gctgcttttc cgctcgttcc tctcctctct gacgtcttct tctgcttctt
     1801 ttctccttcc tttttccctt ctcaatcttc ctctttttcc catttctgtc aatcattcga
     1861 agaagaagaa gaagaagacc ctcattcatt tcattttttt ttctgtcggt gtgtgctgct
     1921 cggttaagag tgagctctct attccacgtc ttcttctttt tcttcttcga ttcgaatcaa
     1981 tcactccacc aaaacgcatt cgtttttggg attcaccccg cggttcgcaa taggtttctt
     2041 tttcaaatta ttagcgttat aaatagaaaa atggtggagt ttcaataaaa atgataattt
     2101 acaaagtgta ttttgattac atgtactcaa aaaggttgaa atttttcata ccagttttcc
     2161 ggaaatccat ctgatatcat tatcgtattt ttctttttaa aaatgttttc aaaaaaaaac
     2221 aaaatatagc tggttatttg gcaccctcta attaccattt tcctgtcaca ccacactctt
     2281 ttccttcttc cacttctttc cgttttcagc cgcttccaac caaaccgata tgaaagccga
     2341 gcaacaacga tcgattccac ccggctcggc gaccttcccg tcgcaggtga gactcagaaa
     2401 actagagaaa ccgctcaact aactcttgat atccgatttc attcttttct cttttctttt
     2461 tgttgaactt ttccatattt ccagatgcca cggccaccac caagcaccga acaaggaatc
     2521 acaacggaat cggagcttgc gaagaaagct caaatcactc cgaacgacgt tttagcactt
     2581 ccgggaatca ctcaaggtat cttgctccct ttggatttca gtttaaaaca taaatttagg
     2641 attcttatgc tccccatctg cgaacatcta taacatcgag ttcaccaagt tccaaatccg
     2701 tgatctggac actgagcaag tgcttttcga gatcgccaaa ccggagaacg atcaggagaa
     2761 tgatgagtcg ccacaggagt cggcaagata cgtgcgttat agatttgctc caaacttttt
     2821 gaaactcaaa acggtcggag caactgtgga attcaaagta ggagacatcc caatccatca
     2881 tttccgaatg atcgaacgtc acttcttcaa agatcgcctt ctgaagtgtt ttgactttga
     2941 attcggattc tgtattccga attcacgaaa caactgtgaa catatctatg agttccctca
     3001 actctctcaa caactcagtg agtcattatt ctaaaaagta caattcaaaa gactaatctc
     3061 tttttcagtg gacgacatga tcaacaatcc aaacgagact cgttctgaca gcttctattt
     3121 cgtcgataac aaactcgtca tgcacaacaa agccgactac tcatatgatg cataaatatt
     3181 taatacaaaa atgttctgga taattattct gtcgaataga aaaaaaactc caaatgtgat
     3241 taaattccaa taattcctgt ctagtttgtt ccttcccctt ccccttctca tgttcaatgc
     3301 attcctaagc ttttcagttc cccccttgtt tctatatttt ttcgctgtcc tgtcacactc
     3361 gctaaaaaca ctaatcacac ggaaatctgt tttcaataaa aactccaact ttaactcatt
     3421 ttcaatttca actgaaagat tttttcatta gagaatgtct agaactaggc cgtttgcctg
     3481 cctatctata acctagtgcc tatcacctac tcgctgcata gtttgatgta cctatataac
     3541 agttcataaa tgaattttgt caaaattcca ctctcaagta ttcttacacg tacttatttt
     3601 gtacaaatcc ttgtcagtcg ctgcaagaac atacacatcg aagaggcgca atcgaaacaa
     3661 caggtgttcg cccgttgatc gtctccccgc cccgcctgtt gggcgggacg ggtgtcggtc
     3721 accaccacca catcatattg ttggacacac tttgcacatc cggttcacac ccgattaccg
     3781 catcggggtc ttgatctggg gcggctattg gatcttttgt ttatgtagat ttttttctcg
     3841 ccgttatgat tttggattat tttttctctt atcttgcagt cgacttgtaa tctttgcctg
     3901 attgctagct agccaaaact ttcacttgtt cttttttttc ctctaagtat tttcccgaac
     3961 ttatagtcga ccttgagctt tgctaaagcc aacaacactt ttcaaatttg cccgcggatc
     4021 cccatctgca gaattcgccc ttaaatgttc actccagaac agttcacaaa agtgatgtcc
     4081 caactgggta acttttccca gttgggacaa atgttccaac ccgatccccg ggattggcca
     4141 aaggacccaa aggtatgttt cgaatgatac taacataaca tagaacattt tcaggaggac
     4201 ccttggaggg taccggtcca cgggctccag gatgacccgg accttcaggc ccttctgaag
     4261 ggcagccagc ttctgaaggt gaagtccagc tcgtggcgta gggaacgctt ctacaagcta
     4321 caggaggact gcaagaccat ctggcaggaa tctcgaaagg tcatgaggtc cccggagtcg
     4381 cagctgttct ccatcgagga cattcaggag gtacggatgg gacaccgcac agaaggcctg
     4441 gagaagtttg cccgagacat ccccgaggat cgatgcttct ccattgtctt caaggaccag
     4501 cgcaacaccc tagacctcat tgccccatca ccagctgacg ctcagcactg ggtgcagggc
     4561 ctgcgcaaga tcatccacca ctccggctcc atggaccagc ggcagaaggg accggtagaa
     4621 aaaatgagta aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat
     4681 ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac
     4741 ggaaaactta cccttaaatt tatttgcact actggaaaac tacctgttcc atgggtaagt
     4801 ttaaacatat atatactaac taaccctgat tatttaaatt ttcagccaac acttgtcact
     4861 actttctgtt atggtgttca atgcttctcg agatacccag atcatatgaa acggcatgac
     4921 tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat
     4981 gacgggaact acaagacacg taagtttaaa cagttcggta ctaactaacc atacatattt
     5041 aaattttcag gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta
     5101 aaaggtattg attttaaaga agatggaaac attcttggac acaaattgga atacaactat
     5161 aactcacaca atgtatacat catggcagac aaacaaaaga atggaatcaa agttgtaagt
     5221 ttaaacatga ttttactaac taactaatct gatttaaatt ttcagaactt caaaattaga
     5281 cacaacattg aagatggaag cgttcaacta gcagaccatt atcaacaaaa tactccaatt
     5341 ggcgatggcc ctgtcctttt accagacaac cattacctgt ccacacaatc tgccctttcg
     5401 aaagatccca acgaaaagag agaccacatg gtccttcttg agtttgtaac agctgctggg
     5461 attacacatg gcatggatga actatacaaa taaggatgat cgacgccaac gtcgttgaat
     5521 tttcaaattt taaatactga atatttgttt tttttcctat tatttattta ttctctttgt
     5581 gttttttttc ttgctttcta aaaaattaat tcaatccaaa tctaaacatt tttttttctc
     5641 tttccgtctc ccaattcgta ttccgctcct ctcatctgaa cacaatgtgc aagtttattt
     5701 atcttctcgc tttcatttca ttaggacgtg gggggaattg gtggaagggg gaaacacaca
     5761 aaaggatgat ggaaatgaaa taaggacaca caatatgcaa caacattcaa ttcagaaata
     5821 tggaggaagg tttaaaagaa aacataaaaa tatatagagg aggaaggaaa actagtaaaa
     5881 aataagcaaa gaaattaggc gaacgatgag aattgtcctc gcttggaaac actggccgtc
     5941 gttttacact cgagacgtac ggtgcgcgcg atgcattcga agatctgccc actagtgagt
     6001 cgtattataa gtgcaagtaa gatcagtgtt tgtttcgata tcagtctgtt tcgtaacggt
     6061 cttctgtata actacaaaaa agaataaaaa accgtatcac aactgagaaa aaagacaaaa
     6121 cttattgaac agtgttcctt tcaaaaaatt attttgtaaa aaatcgagtt tttctccaaa
     6181 ataacgcacg caatcaaaaa caagaaaact aacgtgttct cctgggcgtg gaactcctta
     6241 tcaagattac cattattgtc tgtcagaatt gatatttcaa ttatttcata tctagacaaa
     6301 cccgggtgtc ttgcatttat tagtactgtg agtttacata caaacgaggc ttactcagat
     6361 aacatagagt cacctctaat tagcaaaaac ccatttctaa ttagctattt cgaccagttc
     6421 taaagtcacc gtaattagaa atggtttttg tagcttttct aattagaaat gaccccgaga
     6481 taagtgaaat atggcatttt ctgggttttc tagtaagaaa tgatcccgca cgtagtgatt
     6541 ccgcgttaca agccaaattc aaattaaaaa agtatattac acaattcaag ttacattatt
     6601 ttcaggaaac aatggcgaag agcaaagcag agattcaaaa ggaggaaaga gacaacagat
     6661 atgaacgaag acttcttgaa aagagaaaga gacatgaggt gagtaaccat gataataaat
     6721 aatcaagtaa gggctataga tttataaatt ctggagttaa cagtttcgaa aaaaagtagt
     6781 tttcatatgt gcgaaggaat atcaaaaata ggggaatagc ctaaacgagt gagaaaatta
     6841 aatgtttgtt attttttcaa aaatgtctgc agggccgtta tcctttcaaa tgcactctat
     6901 caaacataga tcatatccat catatcttat ccagaattta tatcatatcc aagattgaga
     6961 gcgtttcaag gaaaatctca aacaaccata caattttgta gatcgacaac aaggcctcca
     7021 cttcttcatc gatggcctct gcagcacctg caccaccaag tttgtatttt tcatattaat
     7081 ataaagtgag ccacgcatag tcgtttttag gatttttgcc agatcgatgt cgcgcaaaca
     7141 gactgttttt ttttctttga aagccgtgcg tgacatgggc cacggattgt aacgccgcgc
     7201 tggatggata aaaatttatt cagtttttta gcctttacaa aatatggttc gactactttc
     7261 tttattttta catttttata ggtgctcatt gagctgagat attttgggta taatcggaaa
     7321 atcgctgatt ttggtcattt ttctgcacta aagattacac aaaaagttca gtatttagat
     7381 tcattgttga aaattgggga tgatctacag atacatatcg ccgtgttcaa aaacacacgt
     7441 gctgtgacgt cgtgactggg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     7501 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     7561 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
     7621 gcggtatttc acaccgcata tggtgcactc tcagtgcaat ctgctctgat gccgcatagt
     7681 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     7741 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     7801 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     7861 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     7921 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     7981 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     8041 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     8101 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     8161 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     8221 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     8281 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     8341 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     8401 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     8461 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     8521 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     8581 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     8641 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     8701 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     8761 cactggggtc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     8821 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     8881 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     8941 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     9001 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     9061 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     9121 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     9181 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     9241 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     9301 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     9361 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     9421 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     9481 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     9541 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     9601 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     9661 cctttttacg gttcctggtc ttttgctggc cttttgctca catgttcttt cctgcgttat
     9721 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc agcggataac
     9781 aatttcacag
//
