LOCUS       sequence-197671-        5400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and mEGFP-linker sequence for N-terminus tagging of 
            human MAP1LC3B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5400)
  AUTHORS   Allen Institute for Cell Science
  TITLE     Allen Institute for Cell Science Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5400)
  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            "Unpublished"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-25:MAP1LC3B-mEGFP
FEATURES             Location/Qualifiers
     source          1..5400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(411..1121,1122..1124,1125..1127))
                     /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(453..474)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     802..821
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1061..1082
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2145..2162)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2145..2161)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2154..2176)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2370..2389)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2489..2511
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2549..2567)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2635..2739
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2740..2808,2809..3600)
                     /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     complement(2958..2977)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3771..4359
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4260..4279
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4513..4530
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4647..4668
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4683..4688,4689..4706,4707..4713)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4721..4737
                     /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)."
     primer_bind     4726..4748
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4745..4761
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4745..4761
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gcggtggccg gggtacggca ggcaccacgc cgcggagggc gcggggtgat tcagcaggcc
       61 cggggccccg agcgcaccgg cccccgcccg ccctccgccg gctgcctcct cccaggcctc
      121 cgaggggtcc gcaggccgcc ccggcccaga ccagcccctg gccctcccgg ctcggccccg
      181 gcctgggtcc ctcacggggc cccgggcccc gcatccctcg gccccgccgg ccggcagcca
      241 ccggccccgc tcggcctccc ggctcggccc cgacccctca cggcgtccca ggccctgccg
      301 ccctccacag ctcggacccc ggccccgccg cacccgccgc cctcgcggcg acactcaccg
      361 aaggtgcgcc gctgcttgaa ggtcttctcc gacgggcttc tcaggccgct cttgtacagc
      421 tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg
      481 cgcttctcgt tggggtcttt gctcagcttg gactgggtgc tcaggtagtg gttgtcgggc
      541 agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg
      601 ctgccgtcct cgatgttgtg gcggatcttg aagttcacct tgatgccgtt cttctgcttg
      661 tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg
      721 ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc
      781 tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc
      841 tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg
      901 gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc
      961 acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg
     1021 ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc
     1081 aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatggt gcggggatct
     1141 gggcggctac ggcggcgacg acgcgagggt cccggcggct cccgggggcg gcggctgctg
     1201 cggcggcgaa tccgactctg gcgatagcca cttcccttgt atctcctcag cccgcagcct
     1261 ggtcaggtga ctcgcgacgc gtcaccgagg ggcggggcca gagccggccg gggccgccgc
     1321 catgacgggc gtggcggaac ccagcagcgg cacgcagcgt gggggcggag caggtgtgtg
     1381 cgcgtgcgcg atgggcgaaa cgcgcgctct gcgcatgctc ccggtcgtgc cccagccccg
     1441 cccagcaccg ctgggcgcct gaggcccctg aggtgacggt tgtgggctgg gccgcacccc
     1501 gggtctgtgg gcgctcggga gcctgctgcc cggccctgag gagatggcag cgcggggcgg
     1561 gggaggctga gccacaggcg gggtgtcccg gggcactctt gagggagggg tcagaggaag
     1621 ggacatgggg caaatccgac catcctttcc cctccagaga gacttggtgg tgctgcccga
     1681 gacgccggca cccggggctg aggcgcgcag ggcaggccgc gccgtctccc ggaggcaggg
     1741 tcgccgctcg cccgtggagc cctggccccc tctgcgtgca ggggcgcgcg tggtccgtgg
     1801 gtggcttccg gggagcggcc gctggtgacc tccgcccgcg gtcactcgac gcccagcctt
     1861 ggcgcgtttg cgcaactgct tttgtcccga gccttcattc tgggcgcagt cccctctccc
     1921 agtccccctg ccgcggcgcc tggaactctc ctggtggctg taagattttc ctaccgttag
     1981 gtcgtctgtg gcgaccgcca ggcctgcccc acatcgctag ccgccctgtc tacccctcag
     2041 cctcccagcc actaaactcg ctggacaacc ttacgctagt gacagttttt gagtctcaga
     2101 ctcatctgtg aaagggcagt cataatccca atacgcgtca attcactggc cgtcgtttta
     2161 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     2221 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     2281 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     2341 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     2401 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     2461 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     2521 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     2581 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     2641 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2701 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2761 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2821 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2881 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2941 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     3001 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3061 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3121 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3181 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3241 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     3301 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3361 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3421 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3481 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3541 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3601 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3661 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3721 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3781 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3841 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3901 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3961 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4021 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4081 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4141 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4201 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4261 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4321 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4381 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4441 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4501 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     4561 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     4621 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     4681 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     4741 cacacaggaa acagctatga ccatgattac gccaagctcg gcgcgccatt gggataccac
     4801 tgtgtgggct cctgacgagg atggcggcag gcattcgcag tccaccgcga tgaacacggt
     4861 gcttcccttc caaaccggtg tctggaactt aggcaacaac agatacctgg atctgcggcg
     4921 ttctcttgct caagcgggaa ataatcactt cgcatcatct gcgtggagta aatattgtga
     4981 tggtgacacc tgcacgttta accaaatgcc agcacaccta agctgggttt ccagtctgaa
     5041 ccggcagaaa agggatgtat tcggtattcc tggtgccaag gtgagaacgt aatcgcaaac
     5101 ccaagagaag tctcccaccc cagctcctgg gctacagaca ggaactggca acaccgctgt
     5161 gggctgtccc gggcgcggcg aggccggggc ccagcacatt cccctcctct cgaccgaggc
     5221 actgaggtct gtaagaggca ctcgcacccg ggcgggctcc ctcgacggga aaaccagccc
     5281 tgaaggtgac gcgctgggtc ccggcgcagc cccgcttcag agccgaacgg cccagcgcgg
     5341 ccccgggggc tgttgggccc cagaagcgcg accctcgccc gcagcgcgct tgcgggggcg
//
