LOCUS       sequence-439880-        5898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology directed repair template for knocking in mGreenLantern 
            reporter to SYCP3..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5898)
  AUTHORS   Pierson Smela M, Adams J, Ma C, Breimann L, Widocki U, Shioda T, 
            Church GM
  TITLE     Induction of Meiosis from Human Pluripotent Stem Cells
  JOURNAL   bioRxiv 2024.05.31.596483
REFERENCE   2  (bases 1 to 5898)
  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: "bioRxiv 
            2024.05.31.596483"
COMMENT     Sequence Label: pHDR2_mGreenLantern-T2A-SYCP3
FEATURES             Location/Qualifiers
     source          1..5898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(125..229)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1305..1307,1308..1310,1311..2021)
                     /codon_start=1
                     /product="monomeric GFP derivative that displays 
                     exceptionally high brightness in cells (Campbell et al., 
                     2020)"
                     /label=mGreenLantern
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLGYGVACFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIVLKGIDFKEDGNILGHKLEYNFNSHKVYITADKQKNGIK
                     ANFKTRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLK
                     ERVTAAGITHDMDELYK"
     primer_bind     complement(1350..1371)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2031..2084
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3406..3978
                     /codon_start=1
                     /gene="Corynebacterium diphtheriae tox"
                     /product="diphtheria toxin fragment A"
                     /label=DTA
                     /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD
                     DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL
                     GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI
                     NFETRGKRGQDAMYEYMAQACAGNRVRR"
     intron          4055..4120
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     rep_origin      complement(4403..4991)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4483..4502)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5162..5898,1..55,56..124))
                     /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     5785..5804
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
       61 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
      121 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
      181 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
      241 gtccgagacc taaattgcgg gacgttttta ttatttgtct tgattattat gagcttattt
      301 ctggcatcct catgtcgtgt cccgccagca actataattt ccccgcgtga ctgagcgccc
      361 tgctagagtt gcccagattt agccagaatc tggcagaggc ggccccaaaa tttggaactc
      421 ccaggcctca ggggccacct ctccagcact gcgcgcttgg agtcggggtg gacactgtca
      481 ttctctctca tcgattgatt gattgagacg aagtctcact ctgtcgccca ggctggagtg
      541 cagtggggcc atctcagctc actgcaacct ccgcctcccg ggttgaagcg attcttctgc
      601 ctcagcctcc tgagtaactg agatcatagg cacccgccac cgcgcccggc taattttgta
      661 tttttagtag agacggggtt tcaccatgtt ggccaggctg gtctcaaact cctgacctca
      721 ggtgatccac ccacctcggc ctcccaaagt gatgggatta caggcatgag ccacagcgcc
      781 cggccatttc tgttgttgtt tttttttttt gagacgaagt ttcactcttg tcacccaggc
      841 tggagtgcag tggctgcgat cttggctcac tgtgacctct gtctcccggg ttcaagcgat
      901 tctcctgcct cagcctcctg agcagctggg actacaggca cgtgccacca cgcccggtta
      961 attttgtttt tttttttcag tagagacggg gtttcatcat gttggccagg ctggtcttga
     1021 actcctgacc tcaggtgatc tgcctgcctc aaccttccaa agtgctggga tttcaggcgt
     1081 gagctaccgt gcttggccta tttattttat tattttttaa caaatgtcat tatcttctta
     1141 acttttggcc ccccaaataa atgaggaaac tgttaaaatt gtattcaaca atccttgttc
     1201 gatatctcct ttgatattta aacttttacc agtgtattaa aatgatgtct ttgcattcag
     1261 aaggttaaag aaatttgttt tttaaagtca ggaagcatct aaatatggtg tccaagggcg
     1321 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     1381 acaagttcag cgtccgcggc gagggcgagg gcgatgccac caacggcaag ctgaccctga
     1441 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttag
     1501 gctacggcgt ggcctgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     1561 agtccgccat gcccgaaggc tacgtccagg agcgcaccat ctctttcaag gacgacggta
     1621 cctacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgtgc
     1681 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     1741 tcaacagcca caaggtctat atcacggccg acaagcagaa gaacggcatc aaggctaact
     1801 tcaagacccg ccacaacgtt gaggacggcg gcgtgcagct cgccgaccac taccagcaga
     1861 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agccatcagt
     1921 ccaaactgag caaagacccc aacgagaagc gcgatcacat ggtcctgaag gagagggtga
     1981 ccgccgccgg gattacacat gacatggacg agctgtacaa ggggtctggc gagggcagag
     2041 gaagtctgct aacatgcggt gacgtcgagg agaatcctgg cccaatggtg tcctccggaa
     2101 aaaagtattc caggaaatct gggaagccgt ctgtggaaga tcagtttacg agagcctatg
     2161 actttgagac tgaagataag aaagatctga gtggatcaga ggaagatgtt attgaaggtg
     2221 cagtacatct catgttgggc agtgatgtcc ttcagttcat ttgttttgaa cccattgaga
     2281 ctatcgtaca acttagtttg taaagcattt tcaggtatgg tcttccctcc cccggtattt
     2341 gtggttacag aacctgctga cacggatggc cagctgtact tgtgtttctc atttgatcct
     2401 tatattgcag tatcggagtt acagaagctt attcggcaac tcttgctttt gcagttattt
     2461 ccaaaatatc tgatatatag tatgtggctt actgtgacat taacccaggg agacttgaaa
     2521 aattattcct ataaaatggt cctgaatcct gatttaattt cacctaacct gcagagaata
     2581 ttgcattttt atgtctgctt tgaaactgta tgttggcaaa aagcctaaaa atccccaaat
     2641 gaagatgttt cattaaaatt taccaaaata acctattcag tttcaaaagc tttttttttt
     2701 tttttaattg tcaatacagg gaagactgca gtcattgaga aacgtaggaa gaaaaggtct
     2761 tctgcaggag tagttgaaga tatggggtaa cgtgttcaaa ataatttttg tttcttagat
     2821 aaagcacttc aagaaaagtt gctaacaaat aatatatgta aaatacttgg catctggtat
     2881 ttcttccaaa acaagcatgg tatgttgtgg atcttctaaa gtgtttaaaa tgtaatttct
     2941 ttccaggggt gaagtgcaga atatgctgga aggagttgga ggtaggtttt aggagatact
     3001 tgtatttaaa gaaaattgtt ttaagccata taatttgtat aataacaaat gttaataatg
     3061 gactataaga ctgttattta atgccttgtt ctcacataga agaacctcca gatcttgtca
     3121 gtgtggtttt caagaggaag caaaaagcct ctccacccag gcctggaatg tttccaccca
     3181 atgtcgagca gtgtggtttt gcaagaggaa gcaaaaagcc tctccaccca ggcctggaat
     3241 gtttccaccc aatgtcgagc aaaccccgcc cagcgtcttg tcattggcga attcgaacac
     3301 gcagatgcag tcggggcggc gcggtcccag gtccacttcg catattaagg tgacgcgtgt
     3361 ggcctcgaac accgagcgac cctgcaggtc ctcgccatgg atcctgatga tgttgttgat
     3421 tcttctaaat cttttgtgat ggaaaacttt tcttcgtacc acgggactaa acctggttat
     3481 gtagattcca ttcaaaaagg tatacaaaag ccaaaatctg gtacacaagg aaattatgac
     3541 gatgattgga aagggtttta tagtaccgac aataaatacg acgctgcggg atactctgta
     3601 gataatgaaa acccgctctc tggaaaagct ggaggcgtgg tcaaagtgac gtatccagga
     3661 ctgacgaagg ttctcgcact aaaagtggat aatgccgaaa ctattaagaa agagttaggt
     3721 ttaagtctca ctgaaccgtt gatggagcaa gtcggaacgg aagagtttat caaaaggttc
     3781 ggtgatggtg cttcgcgtgt agtgctcagc cttcccttcg ctgaggggag ttctagcgtt
     3841 gaatatatta ataactggga acaggcgaaa gcgttaagcg tagaacttga gattaatttt
     3901 gaaacccgtg gaaaacgtgg ccaagatgcg atgtatgagt atatggctca agcctgtgca
     3961 ggaaatcgtg tcaggcgatc tctttgtgaa ggaaccttac ttctgtggtg tgacataatt
     4021 ggacaaacta cctacagaga tttaaagctc taaggtaaat ataaaatttt taagtgtata
     4081 atgtgttaaa ctactgattc taattgtttg tgtattttag attccaacct atggaactga
     4141 tgaatgggag cagtggtgga atgcagatcc actagagtcg agggatgcag aaattgatga
     4201 tctattaaac aataaagatg tccactaaaa tggaagtttt tcctgtcata ctttgttaag
     4261 aagggtgaga acagagtacc tacattttga atggaaggat tggagctacg ggggtggggg
     4321 tggggtggga ttagataaat gcctgctctt tactgaaggc tctttactat tgctttatga
     4381 taatgtttca tagttggatg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4441 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4501 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4561 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4621 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4681 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4741 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4801 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4861 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4921 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4981 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5041 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5101 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5161 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5221 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5281 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5341 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5401 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5461 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5521 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5581 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5641 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5701 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5761 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5821 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5881 ccagttcgat gtaaccca
//
