LOCUS       sequence-330361-        5406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and mEGFP-linker sequence for N-terminus tagging of 
            human TERF2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5406)
  TITLE     Allen Institute for Cell Science Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5406)
  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: 
            "Unpublished"
COMMENT     Sequence Label: AICSDP-77:TERF2-mEGFP
FEATURES             Location/Qualifiers
     source          1..5406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..80
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     72..89
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     73..89
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1107..1109,1110..1112,1113..1823)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1152..1173)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1413..1432)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1760..1781
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2879..2895)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2879..2895)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2892..2914)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2903..2919
                     /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(2927..2933,2934..2951,2952..2957))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2972..2993
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3110..3127)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3281..3869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3361..3380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4040..4831,4832..4900))
                     /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     4663..4682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4901..5005)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5073..5091
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5129..5151)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5251..5270
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
       61 agtcacgacg ttgtaaaacg acggccagtg aattgacgcg tattgggata actggatctt
      121 tgcaggttcc agactagtgg ggctggtagg acaatctttg aagctagacc agttactggg
      181 cctctcagga agaggcaccc acaaccacca tcctgaacta aggtctgagc tggaggttct
      241 ccatttggtc aagtgccagt tggtgaattc caagagtatc agagatatct gaaagtgact
      301 cagaccctct atgggactat gggtggtcct tttgcttatt tttctaattg ttctacaaca
      361 aaatgtgtta tttttgtgat tacaaagagt atatgtctag cggctgggtt gccaggtgca
      421 ttctatttat tggtatcaca ctggtctttg ctttataatt acagtatttg ttaaaataca
      481 aatttgttta aagactttcc tctgtgtatg ctctcttcca caattttcaa agtgtttaat
      541 aaggtgcaca tatttggttt tctttttttt cttttctttt ttttgagacg gagtttcgct
      601 cttgttgccc agactggagt gcaatggcgc gatctcggct caccgcaacc tccgcctcgc
      661 gggttcaagc gagtctcctg cctcagcctc ctgagtagct aggattacag gcatgcgcca
      721 ccacgtctgg ctaattttgt atttttagta gagatggggt ttctccatgt tggtcaggct
      781 ggtctcgaac tcccgacctc aggtgatcca cccgcctcag cctcccaaag tgctgggatt
      841 acaggcgtag agccaccgat cccggcctgt ttttcagtct ctaatacact gttccgcgtc
      901 cctagcatag tagctgtttt ctgtaaattc ggagcgaagt tgactgcaaa acccgccggg
      961 ttcaccggcc tgggcggtgt ggacttcaag ccccaggagg cattgcggcc ggcacatcgg
     1021 gaactacggc gtctgagaag ggggccgggc cctccggtcc tttccctccc agaagcccgc
     1081 gggcggctcg gaacgctgtt tctatcatgg tgagcaaggg cgaggagctg ttcaccgggg
     1141 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     1201 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     1261 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     1321 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1381 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1441 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1501 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     1561 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     1621 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1681 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc
     1741 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1801 tcggcatgga cgagctgtac aagagcggca gaacccagat ccccgcagcg ggagcaggga
     1861 cggcgggccc cgcttccggc ccgggcgtcg tgcgtgaccc agcggcgtca cagccgagga
     1921 agcggcccgg ccgggagggc ggggagggcg cgcggcgatc ggacacgatg gcgggaggag
     1981 gcgggagtag cgacggcagc gggcgggcag ctggcaggcg ggcgtcccgc agtagcgggc
     2041 gggcccggcg ggggcgccac gagccggggc tggggggccc ggcggagcgc ggcgcggggg
     2101 aggcacggct ggaagaggca gtcaatcgct gggtgctcaa gttctacttc cacgaggcgc
     2161 tgcgggcctt tcggggtagc cggtacgggg acttcagaca gatccgggac atcatgcagg
     2221 gtgagggccg ggccggggga ggggggttgg agcgccgggg aagggggtcc gggggaccgg
     2281 gagtcgggac tgggggtcgc ccagccaacg atgtcttctc tcccagcttt gcttgtcagg
     2341 cccttgggga aggagcacac cgtgtcccga ttgctgcggg ttatgcagtg tctgtcgcgg
     2401 attgaagaag gggaaaattt aggtatggct atttttgtgt tcttctctgg gcttacttgc
     2461 gtagggtttt gttttaaaac ttatatctag aactgaagga ttccaggtta aagtgcgata
     2521 gtttccatca gagtggtctt tttcattcta cgatgggtct gttttctggc tcgactggct
     2581 ctagttttct ccggttaagg ttgtcttggc tgcttttctg tgtttttttt aaatcttctc
     2641 ccacctgtta attgctcacc ccaactttgc tcctgtctaa agtgagttat gttcatatgg
     2701 acatcttttt tcccccatgg tagagagttt ttcagagtgg aagtactctg acatttttcc
     2761 cctctgaatt taacttgaaa gctatttagc tgttgtcagt ttcttgccac tcgtatctga
     2821 ggaggatggg tatattctga atttatccca atggcgcgcc gagcttggcg taatcatggt
     2881 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     2941 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     3001 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3061 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3121 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3181 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3241 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3301 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3361 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3421 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3481 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3541 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3601 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3661 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3721 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3781 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3841 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3901 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3961 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4021 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4081 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4141 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4201 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4261 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4321 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4381 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4441 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4501 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4561 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4621 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4681 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4741 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4801 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4861 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4921 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4981 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     5041 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5101 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     5161 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     5221 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     5281 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     5341 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     5401 tacgcc
//
