LOCUS       sequence-210698-       10291 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP reporter of lag-2 driven by 1 kb promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10291)
  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 10291)
  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: pGC642
FEATURES             Location/Qualifiers
     source          1..10291
                     /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(5155..5183)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5931..5951
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(6432..6449)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6432..6448)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8275..8297
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8335..8353)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8421..8525
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8526..8594,8595..9386)
                     /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(8744..8763)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9557..10145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10046..10065
                     /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 cgttttaaat
     3481 tagttttcga attcccgctc aaataaatgt cccagaattt acatttttga gttttccggc
     3541 taaatttttg ccgttttttg aatatgattt gtaatatttg gtctgaacca tggtttttcc
     3601 aattttcaga atctttctcg caaaaaatct ccgtatcccg caaaataatt gttgaagcgt
     3661 caaaaattgg cctaaaagtc gataaattaa cattgatttc gctttttttt taggaaaaaa
     3721 gtctaaaagt attttttaaa ctatacaaaa tcattaaatt ttgacaagaa aacgcttaaa
     3781 ttacaatttt ttttattttt taaaaattta aaaaaatttt aaaaaaacat taaaattcag
     3841 ataaattgca caatttttat tgcacaagtc gttcgtcgtt cgtgtttttt tcgctgtcaa
     3901 aatgttggca aagattgtga agtcccctgt agtttaacac tctaagttac tccaaagact
     3961 tctacctgcc tacctgcctg cctatctata acctagtgcc tatcacctac tcgctgcata
     4021 gtttgatgta cctatataac agttcataaa tgaattttgt caaaattcca ctctcaagta
     4081 ttcttacacg tacttatttt gtacaaatcc ttgtcagtcg ctgcaagaac atacacatcg
     4141 aagaggcgca atcgaaacaa caggtgttcg cccgttgatc gtctccccgc cccgcctgtt
     4201 gggcgggacg ggtgtcggtc accaccacca catcatattg ttggacacac tttgcacatc
     4261 cggttcacac ccgattaccg catcggggtc ttgatctggg gcggctattg gatcttttgt
     4321 ttatgtagat ttttttctcg ccgttatgat tttggattat tttttctctt atcttgcagt
     4381 cgacttgtaa tctttgcctg attgctagct agccaaaact ttcacttgtt cttttttttc
     4441 ctctaagtat tttcccgaac ttatagtcga ccttgagctt tgctagaagc caataacact
     4501 tttcaaattt gcccgcggat ccccatctgc agaattcgcc cttaaatgtt cactccagaa
     4561 cagttcacaa aagtgatgtc ccaactgggt aacttttccc agttgggaca aatgttccaa
     4621 cccgatcccc gggattggcc aaaggaccca aaggtatgtt tcgaatgata ctaacataac
     4681 atagaacatt ttcaggagga cccttggagg gtaccggtcc acgggctcca ggatgacccg
     4741 gaccttcagg cccttctgaa gggcagccag cttctgaagg tgaagtccag ctcgtggcgt
     4801 agggaacgct tctacaagct acaggaggac tgcaagacca tctggcagga atctcgaaag
     4861 gtcatgaggt ccccggagtc gcagctgttc tccatcgagg acattcagga ggtacggatg
     4921 ggacaccgca cagaaggcct ggagaagttt gcccgagaca tccccgagga tcgatgcttc
     4981 tccattgtct tcaaggacca gcgcaacacc ctagacctca ttgccccatc accagctgac
     5041 gctcagcact gggtgcaggg cctgcgcaag atcatccacc actccggctc catggaccag
     5101 cggcagaagg gaccggtaga aaaaatgagt aaaggagaag aacttttcac tggagttgtc
     5161 ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag
     5221 ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac tactggaaaa
     5281 ctacctgttc catgggtaag tttaaacata tatatactaa ctaaccctga ttatttaaat
     5341 tttcagccaa cacttgtcac tactttctgt tatggtgttc aatgcttctc gagataccca
     5401 gatcatatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa
     5461 agaactatat ttttcaaaga tgacgggaac tacaagacac gtaagtttaa acagttcggt
     5521 actaactaac catacatatt taaattttca ggtgctgaag tcaagtttga aggtgatacc
     5581 cttgttaata gaatcgagtt aaaaggtatt gattttaaag aagatggaaa cattcttgga
     5641 cacaaattgg aatacaacta taactcacac aatgtataca tcatggcaga caaacaaaag
     5701 aatggaatca aagttgtaag tttaaacatg attttactaa ctaactaatc tgatttaaat
     5761 tttcagaact tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat
     5821 tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg
     5881 tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt
     5941 gagtttgtaa cagctgctgg gattacacat ggcatggatg aactatacaa ataaggatga
     6001 tcgacgccaa cgtcgttgaa ttttcaaatt ttaaatactg aatatttgtt ttttttccta
     6061 ttatttattt attctctttg tgtttttttt cttgctttct aaaaaattaa ttcaatccaa
     6121 atctaaacat ttttttttct ctttccgtct cccaattcgt attccgctcc tctcatctga
     6181 acacaatgtg caagtttatt tatcttctcg ctttcatttc attaggacgt ggggggaatt
     6241 ggtggaaggg ggaaacacac aaaaggatga tggaaatgaa ataaggacac acaatatgca
     6301 acaacattca attcagaaat atggaggaag gtttaaaaga aaacataaaa atatatagag
     6361 gaggaaggaa aactagtaaa aaataagcaa agaaattagg cgaacgatga gaattgtcct
     6421 cgcttggaaa cactggccgt cgttttacac tcgagacgta cggtgcgcgc gatgcattcg
     6481 aagatctgcc cactagtgag tcgtattata agtgcaagta agatcagtgt ttgtttcgat
     6541 atcagtctgt ttcgtaacgg tcttctgtat aactacaaaa aagaataaaa aaccgtatca
     6601 caactgagaa aaaagacaaa acttattgaa cagtgttcct ttcaaaaaat tattttgtaa
     6661 aaaatcgagt ttttctccaa aataacgcac gcaatcaaaa acaagaaaac taacgtgttc
     6721 tcctgggcgt ggaactcctt atcaagatta ccattattgt ctgtcagaat tgatatttca
     6781 attatttcat atctagacaa acccgggtgt cttgcattta ttagtactgt gagtttacat
     6841 acaaacgagg cttactcaga taacatagag tcacctctaa ttagcaaaaa cccatttcta
     6901 attagctatt tcgaccagtt ctaaagtcac cgtaattaga aatggttttt gtagcttttc
     6961 taattagaaa tgaccccgag ataagtgaaa tatggcattt tctgggtttt ctagtaagaa
     7021 atgatcccgc acgtagtgat tccgcgttac aagccaaatt caaattaaaa aagtatatta
     7081 cacaattcaa gttacattat tttcaggaaa caatggcgaa gagcaaagca gagattcaaa
     7141 aggaggaaag agacaacaga tatgaacgaa gacttcttga aaagagaaag agacatgagg
     7201 tgagtaacca tgataataaa taatcaagta agggctatag atttataaat tctggagtta
     7261 acagtttcga aaaaaagtag ttttcatatg tgcgaaggaa tatcaaaaat aggggaatag
     7321 cctaaacgag tgagaaaatt aaatgtttgt tattttttca aaaatgtctg cagggccgtt
     7381 atcctttcaa atgcactcta tcaaacatag atcatatcca tcatatctta tccagaattt
     7441 atatcatatc caagattgag agcgtttcaa ggaaaatctc aaacaaccat acaattttgt
     7501 agatcgacaa caaggcctcc acttcttcat cgatggcctc tgcagcacct gcaccaccaa
     7561 gtttgtattt ttcatattaa tataaagtga gccacgcata gtcgttttta ggatttttgc
     7621 cagatcgatg tcgcgcaaac agactgtttt tttttctttg aaagccgtgc gtgacatggg
     7681 ccacggattg taacgccgcg ctggatggat aaaaatttat tcagtttttt agcctttaca
     7741 aaatatggtt cgactacttt ctttattttt acatttttat aggtgctcat tgagctgaga
     7801 tattttgggt ataatcggaa aatcgctgat tttggtcatt tttctgcact aaagattaca
     7861 caaaaagttc agtatttaga ttcattgttg aaaattgggg atgatctaca gatacatatc
     7921 gccgtgttca aaaacacacg tgctgtgacg tcgtgactgg ggaaaaccct ggcgttaccc
     7981 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     8041 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     8101 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtgcaa
     8161 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     8221 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     8281 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     8341 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     8401 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     8461 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     8521 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     8581 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     8641 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     8701 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     8761 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     8821 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     8881 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     8941 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     9001 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     9061 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     9121 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     9181 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     9241 ggtctcgcgg tatcattgca gcactggggt cagatggtaa gccctcccgt atcgtagtta
     9301 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     9361 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     9421 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     9481 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     9541 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     9601 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     9661 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     9721 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     9781 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     9841 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     9901 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     9961 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
    10021 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
    10081 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
    10141 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggt cttttgctgg ccttttgctc
    10201 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
    10261 gagctgatac cagcggataa caatttcaca g
//
