LOCUS       sequence-454212-        5808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and mEGFP sequence for C-terminus tagging of human 
            cadherin 5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5808)
  AUTHORS   Allen Institute
  TITLE     Allen Institute for Cell Science Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5808)
  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            "Unpublished"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-167:CDH5-mEGFP
FEATURES             Location/Qualifiers
     source          1..5808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   1007..1147
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1018..1147
                     /label=AAV2 ITR
     CDS             complement(1674..1685)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(1983..1985,1986..2699)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2025..2046)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2286..2305)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2633..2654
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     repeat_region   3557..3697
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3557..3686
                     /label=AAV2 ITR
     primer_bind     complement(3707..3724)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3707..3723)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3716..3738)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3936..4391
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4023..4042)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4233..4254
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4408..4427)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4527..4549
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4587..4605)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4673..4777
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4778..4846,4847..5638)
                     /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(4996..5015)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gaattgcctg caggcagctg
     1021 cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt
     1081 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
     1141 ggttcctatc gatggcgcgc ctccagatat catacactcg agtgggagtg tcgggcgagg
     1201 tggggatgag tatgcggggg agcggcaggg acggggtgtg gggtgtaggg ggatggcggg
     1261 gatgggggta tgggggaacg gcggggatgg ggttgcaggg gaaggggggc caaaaggatg
     1321 gggttgcagg ggaagggggg gccaaaggga tgggcatgct gggagggcgt gggggtgggg
     1381 ttgcaggggg cagtggggat gggggcatgg ggggccgccc aggcccccag ccttgtctga
     1441 ctctgctgct cggctccctg gctgcagtga tcaccctgct catcttcctg cggcggcggc
     1501 tccggaagca ggcccgcgcg cacggcaaga gcgtgccgga gatccacgag cagctggtca
     1561 cctacgacga ggagggcggc ggcgagatgg acaccaccag ctacgatgtg tcggtgctca
     1621 actcggtgcg ccgcggcggg gccaagcccc cgcggcccgc gctggacgcc cggccttccc
     1681 tctatgcgca ggtgcagaag ccaccgaggc acgcgcctgg ggcacacgga gggcccgggg
     1741 agatggcagc catgatcgag gtgaagaagg acgaggcgga ccacgacggc gacggccccc
     1801 cctacgacac gctgcacatc tacggctacg agggctccga gtccatagcc gagtccctca
     1861 gctccctggg caccgactca tccgactctg acgtggatta cgacttcctt aacgactggg
     1921 gacccaggtt taagatgctg gctgagctgt acggctcgga cccccgggag gagctgctgt
     1981 atgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2041 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     2101 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2161 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     2221 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     2281 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2341 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2401 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     2461 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     2521 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2581 acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2641 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat
     2701 aggcggccga ggtcactctg tgcctgggga cccaaacccc ctgcagccca ggccagtcag
     2761 acgccaggca ccacagcctc caaaaatggc agtgactccc cagcccagca ccccttcctc
     2821 gtgggtccca gagacctcat cagccttggg atagcaaact ccaggttcct gaaatatcca
     2881 ggaatatatg tcagtgatga ctattctcaa atgctggcaa atccaggctg gtgttctgtc
     2941 tgggctcaga catccacata accctgtcac ccacagaccg ccgtctaact caaagacttc
     3001 ctctggctcc ccaaggctgc aaagcaaaac agactgtgtt taactgctgc agggtctttt
     3061 tctagggtcc ctgaacgccc tggtaaggct ggtgaggtcc tggtgcctat ctgcctggag
     3121 gcaaaggcct ggacagcttg acttgtgggg caggattctc tgcagcccat tcccaaggga
     3181 gactgaccat catgccctct ctcgggagcc ctagccctgc tccaactcca tactccactc
     3241 caagtgcccc accactcccc aacccctctc caggcctgtc aagagggagg aaggggcccc
     3301 atggcagctc ctgaccttgg gtcctgaagt gacctcactg gcctgccatg ccagtaactg
     3361 tgctgtactg agcactgaac cacattcagg gaaatggctt attaaacttt gaagcaactg
     3421 tgaattcatt ctggaggggc agtggagatc aggagtgaca gatcacaggg tgagggccac
     3481 ctccacaccc accccctctg gccggccgct tcgagttaat taatccaacc ggttaccgcc
     3541 taggatcgat agatctagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3601 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3661 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggcag cttggcactg gccgtcgttt
     3721 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     3781 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     3841 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     3901 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     3961 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4021 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4081 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4141 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4201 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4261 actcaaccct atctcgggct attcttttga tttataaggg attttgccga tttcggccta
     4321 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4381 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4441 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4501 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4561 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4621 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4681 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4741 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4801 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4861 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4921 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4981 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5041 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5101 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5161 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5221 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5281 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5341 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5401 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5461 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5521 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5581 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5641 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5701 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5761 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
