LOCUS       sequence-210687-       11291 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP
            of lag-2 driven by 2 kb promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11291)
  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 11291)
  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: pGC630
FEATURES             Location/Qualifiers
     source          1..11291
                     /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(6155..6183)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     6931..6951
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(7432..7449)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7432..7448)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9275..9297
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9335..9353)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9421..9525
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9526..9594,9595..10386)
                     /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(9744..9763)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10557..11145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11046..11065
                     /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 cgttttttcg
     3481 gaacgtctca ttacaaaatt tggttgcttt gagccatttt gaatccgaaa aagtctcaaa
     3541 aagcaaagtc ttaaatttcg gtactccacc tttaaaaaaa atagggtttc cgccgaaatt
     3601 tctgggttcc aataatatct gttaccattt ccttgctctg cgacgagcag ccaccagaca
     3661 aaaatctcct aatacccctc ttcatgcctt tgttcttccg gtttggcggt ctgaaaaaga
     3721 gcacccctcc cggcgggggg ctctcggtat ctgttccgca aaaaccgtca tatggagggc
     3781 tatcgggaga cgggaagagt tggaaatacg ggaaatggtg atgaataggt agatcaatcg
     3841 attccggaac aaataatttg agttgagcac gcgtgctcca catgtgctca gccgcttagg
     3901 tgtgagtgaa gatccttgga tttcaaggcc atcagccatt tcccctgtcc agctagagca
     3961 gtcctaaacg gaattttttg cattcttttg cttctttaca atttcaaatg ttcactctaa
     4021 ctaattcaaa aatctttctt ttttacgagc tctccttgct cgccgttgta ttccattcat
     4081 acctttaaga ccacccgagt gtacgcatat ttaaccaggc accctaaaaa catgaatagt
     4141 ctgtggtgga ggtagacagt gtgttttcta agaatgtctc cgtaatttga aatatatttt
     4201 agagtcacct gaaattcatt tttcaagcaa tatttcttac catatctttg gggataattt
     4261 taaaagtgga gtgaagtcgt ttagggacac gcttttaagt tttaggaaat ggtttttttt
     4321 taattcataa cttaaaaaaa tctataaaaa ttgttacatt ttttatgatt ttctcaattt
     4381 tgttgaatgt ttgaattttt tttttgagat gtattgtttg aattggagct tttttgtgag
     4441 ttttttttta attttgcaat taaataaaaa gtagctaaat tagttttcga attcccgctc
     4501 aaataaatgt cccagaattt acatttttga gttttccggc taaatttttg ccgttttttg
     4561 aatatgattt gtaatatttg gtctgaacca tggtttttcc aattttcaga atctttctcg
     4621 caaaaaatct ccgtatcccg caaaataatt gttgaagcgt caaaaattgg cctaaaagtc
     4681 gataaattaa cattgatttc gctttttttt taggaaaaaa gtctaaaagt attttttaaa
     4741 ctatacaaaa tcattaaatt ttgacaagaa aacgcttaaa ttacaatttt ttttattttt
     4801 taaaaattta aaaaaatttt aaaaaaacat taaaattcag ataaattgca caatttttat
     4861 tgcacaagtc gttcgtcgtt cgtgtttttt tcgctgtcaa aatgttggca aagattgtga
     4921 agtcccctgt agtttaacac tctaagttac tccaaagact tctacctgcc tacctgcctg
     4981 cctatctata acctagtgcc tatcacctac tcgctgcata gtttgatgta cctatataac
     5041 agttcataaa tgaattttgt caaaattcca ctctcaagta ttcttacacg tacttatttt
     5101 gtacaaatcc ttgtcagtcg ctgcaagaac atacacatcg aagaggcgca atcgaaacaa
     5161 caggtgttcg cccgttgatc gtctccccgc cccgcctgtt gggcgggacg ggtgtcggtc
     5221 accaccacca catcatattg ttggacacac tttgcacatc cggttcacac ccgattaccg
     5281 catcggggtc ttgatctggg gcggctattg gatcttttgt ttatgtagat ttttttctcg
     5341 ccgttatgat tttggattat tttttctctt atcttgcagt cgacttgtaa tctttgcctg
     5401 attgctagct agccaaaact ttcacttgtt cttttttttc ctctaagtat tttcccgaac
     5461 ttatagtcga ccttgagctt tgctagaagc caataacact tttcaaattt gcccgcggat
     5521 ccccatctgc agaattcgcc cttaaatgtt cactccagaa cagttcacaa aagtgatgtc
     5581 ccaactgggt aacttttccc agttgggaca aatgttccaa cccgatcccc gggattggcc
     5641 aaaggaccca aaggtatgtt tcgaatgata ctaacataac atagaacatt ttcaggagga
     5701 cccttggagg gtaccggtcc acgggctcca ggatgacccg gaccttcagg cccttctgaa
     5761 gggcagccag cttctgaagg tgaagtccag ctcgtggcgt agggaacgct tctacaagct
     5821 acaggaggac tgcaagacca tctggcagga atctcgaaag gtcatgaggt ccccggagtc
     5881 gcagctgttc tccatcgagg acattcagga ggtacggatg ggacaccgca cagaaggcct
     5941 ggagaagttt gcccgagaca tccccgagga tcgatgcttc tccattgtct tcaaggacca
     6001 gcgcaacacc ctagacctca ttgccccatc accagctgac gctcagcact gggtgcaggg
     6061 cctgcgcaag atcatccacc actccggctc catggaccag cggcagaagg gaccggtaga
     6121 aaaaatgagt aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga
     6181 tggtgatgtt aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata
     6241 cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc catgggtaag
     6301 tttaaacata tatatactaa ctaaccctga ttatttaaat tttcagccaa cacttgtcac
     6361 tactttctgt tatggtgttc aatgcttctc gagataccca gatcatatga aacggcatga
     6421 ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa agaactatat ttttcaaaga
     6481 tgacgggaac tacaagacac gtaagtttaa acagttcggt actaactaac catacatatt
     6541 taaattttca ggtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt
     6601 aaaaggtatt gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta
     6661 taactcacac aatgtataca tcatggcaga caaacaaaag aatggaatca aagttgtaag
     6721 tttaaacatg attttactaa ctaactaatc tgatttaaat tttcagaact tcaaaattag
     6781 acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa atactccaat
     6841 tggcgatggc cctgtccttt taccagacaa ccattacctg tccacacaat ctgccctttc
     6901 gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg
     6961 gattacacat ggcatggatg aactatacaa ataaggatga tcgacgccaa cgtcgttgaa
     7021 ttttcaaatt ttaaatactg aatatttgtt ttttttccta ttatttattt attctctttg
     7081 tgtttttttt cttgctttct aaaaaattaa ttcaatccaa atctaaacat ttttttttct
     7141 ctttccgtct cccaattcgt attccgctcc tctcatctga acacaatgtg caagtttatt
     7201 tatcttctcg ctttcatttc attaggacgt ggggggaatt ggtggaaggg ggaaacacac
     7261 aaaaggatga tggaaatgaa ataaggacac acaatatgca acaacattca attcagaaat
     7321 atggaggaag gtttaaaaga aaacataaaa atatatagag gaggaaggaa aactagtaaa
     7381 aaataagcaa agaaattagg cgaacgatga gaattgtcct cgcttggaaa cactggccgt
     7441 cgttttacac tcgagacgta cggtgcgcgc gatgcattcg aagatctgcc cactagtgag
     7501 tcgtattata agtgcaagta agatcagtgt ttgtttcgat atcagtctgt ttcgtaacgg
     7561 tcttctgtat aactacaaaa aagaataaaa aaccgtatca caactgagaa aaaagacaaa
     7621 acttattgaa cagtgttcct ttcaaaaaat tattttgtaa aaaatcgagt ttttctccaa
     7681 aataacgcac gcaatcaaaa acaagaaaac taacgtgttc tcctgggcgt ggaactcctt
     7741 atcaagatta ccattattgt ctgtcagaat tgatatttca attatttcat atctagacaa
     7801 acccgggtgt cttgcattta ttagtactgt gagtttacat acaaacgagg cttactcaga
     7861 taacatagag tcacctctaa ttagcaaaaa cccatttcta attagctatt tcgaccagtt
     7921 ctaaagtcac cgtaattaga aatggttttt gtagcttttc taattagaaa tgaccccgag
     7981 ataagtgaaa tatggcattt tctgggtttt ctagtaagaa atgatcccgc acgtagtgat
     8041 tccgcgttac aagccaaatt caaattaaaa aagtatatta cacaattcaa gttacattat
     8101 tttcaggaaa caatggcgaa gagcaaagca gagattcaaa aggaggaaag agacaacaga
     8161 tatgaacgaa gacttcttga aaagagaaag agacatgagg tgagtaacca tgataataaa
     8221 taatcaagta agggctatag atttataaat tctggagtta acagtttcga aaaaaagtag
     8281 ttttcatatg tgcgaaggaa tatcaaaaat aggggaatag cctaaacgag tgagaaaatt
     8341 aaatgtttgt tattttttca aaaatgtctg cagggccgtt atcctttcaa atgcactcta
     8401 tcaaacatag atcatatcca tcatatctta tccagaattt atatcatatc caagattgag
     8461 agcgtttcaa ggaaaatctc aaacaaccat acaattttgt agatcgacaa caaggcctcc
     8521 acttcttcat cgatggcctc tgcagcacct gcaccaccaa gtttgtattt ttcatattaa
     8581 tataaagtga gccacgcata gtcgttttta ggatttttgc cagatcgatg tcgcgcaaac
     8641 agactgtttt tttttctttg aaagccgtgc gtgacatggg ccacggattg taacgccgcg
     8701 ctggatggat aaaaatttat tcagtttttt agcctttaca aaatatggtt cgactacttt
     8761 ctttattttt acatttttat aggtgctcat tgagctgaga tattttgggt ataatcggaa
     8821 aatcgctgat tttggtcatt tttctgcact aaagattaca caaaaagttc agtatttaga
     8881 ttcattgttg aaaattgggg atgatctaca gatacatatc gccgtgttca aaaacacacg
     8941 tgctgtgacg tcgtgactgg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     9001 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     9061 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     9121 tgcggtattt cacaccgcat atggtgcact ctcagtgcaa tctgctctga tgccgcatag
     9181 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     9241 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     9301 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     9361 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     9421 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     9481 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     9541 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     9601 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     9661 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     9721 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     9781 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     9841 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     9901 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     9961 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
    10021 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
    10081 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
    10141 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
    10201 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
    10261 gcactggggt cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
    10321 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
    10381 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
    10441 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
    10501 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
    10561 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
    10621 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
    10681 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
    10741 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
    10801 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
    10861 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
    10921 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
    10981 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
    11041 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
    11101 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
    11161 gcctttttac ggttcctggt cttttgctgg ccttttgctc acatgttctt tcctgcgtta
    11221 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cagcggataa
    11281 caatttcaca g
//
