LOCUS       sequence-252486-        4966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes TU with connectors L2 R3 and an C3 insulator, and pCAG 
            expressing membrane-tagged mAzamiGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4966)
  AUTHORS   Fonseca JP, Bonny AR, Kumar GR, Ng AH, Town J, Wu QC, Aslankoohi E, 
            Chen SY, Dods G, Harrigan P, Osimiri LC, Kistler AL, El-Samad H
  TITLE     A Toolkit for Rapid Modular Construction of Biological Circuits in 
            Mammalian Cells
  JOURNAL   ACS Synth Biol. 2019 Nov 15;8(11):2593-2606. doi: 
            10.1021/acssynbio.9b00322. Epub 2019 Nov 5.
  PUBMED    31686495
REFERENCE   2  (bases 1 to 4966)
  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: 
            "10.1021/acssynbio.9b00322"; journalName: "ACS Synth Biol"; date: 
            "2019-11-15- 15"; volume: "8"; issue: "11"; pages: "2593-2606"
COMMENT     Sequence Label: ARB350
FEATURES             Location/Qualifiers
     source          1..4966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(103..894,895..963))
                     /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     726..745
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(964..1068)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1171..1759
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1660..1679
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2269..2572
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2574..2850
                     /label=chicken beta-actin promoter
     intron          2851..3867
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3791..3813
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3875..3894
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3973..4644
                     /codon_start=1
                     /product="humanized monomeric Azami-Green fluorescent 
                     protein"
                     /label=hmAzami-Green
                     /note="mammalian codon-optimized"
                     /translation="VSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNVT
                     EGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTAT
                     SNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLLL
                     EGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLPS
                     QA"
     CDS             3973..4644
                     /codon_start=1
                     /product="monomeric Azami-Green fluorescent protein 
                     (Karasawa et al., 2003)"
                     /label=mAzami-Green
                     /translation="VSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNVT
                     EGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTAT
                     SNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLLL
                     EGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLPS
                     QA"
     primer_bind     complement(4684..4703)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4738..4757
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 gcggccgcga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
       61 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
      121 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
      181 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
      241 gcgggaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
      301 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
      361 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
      421 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
      481 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
      541 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
      601 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
      661 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
      721 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
      781 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
      841 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
      901 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
      961 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1021 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1081 aaaagtgcca cctgtcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1141 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1201 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1261 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1321 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1381 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1441 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1501 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1561 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1621 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     1681 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     1741 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     1801 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtgc
     1861 ggccgcccct gaattcgcat ctagaataaa tcgcagccaa gtgagtgaat agatgacgca
     1921 ccacggtcag acacggccac atcgtatctc acaggagcaa gcgcgatagg agcactcaca
     1981 catagtacgg tgatccgctg actcctttgc ccaaataaga cgtgagcccg tctcagatgc
     2041 acacactggc ttaagatgac gggaccctgt aaacgtgtcc gccttgccac tttgtgcaat
     2101 atttagctcc ttcagtagag ggcgctgcag gaacatgtca ggagggggcc ttctcttccg
     2161 gattctggat tctggtatac acttggagcg tggcatttgg gggacagcca ctggtgcgca
     2221 gccccatcaa gttttggtgg catccctaac gccatatatg gagttccgcg ttacataact
     2281 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     2341 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     2401 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     2461 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     2521 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt
     2581 gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta
     2641 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg gnngcgcgcg
     2701 ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca
     2761 gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg
     2821 ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg
     2881 ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
     2941 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga
     3001 cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg
     3061 tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc
     3121 ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc
     3181 cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc
     3241 gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc
     3301 gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc gagttgctga gcacggcccg
     3361 gcttcgggtg cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg
     3421 tggcggcagg tgggggtgcc gggcggggcg gggccgcctc gggccgggga gggctcgggg
     3481 gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat
     3541 tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg
     3601 gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc
     3661 ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc
     3721 cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg
     3781 ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat
     3841 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct
     3901 catcattttg gcaaagaatt cgccactatg ggctgtatca agagcaagag gaaggacgtc
     3961 gagaacggtt ctgtgagcgt gatcaagccc gagatgaaga tcaagctgtg catgaggggc
     4021 accgtgaacg gccacaactt cgtgatcgag ggcgagggca agggcaaccc ctacgagggc
     4081 acccagatcc tggacctgaa cgtgaccgag ggcgcccccc tgcccttcgc ctacgacatc
     4141 ctgaccaccg tgttccagta cggcaacagg gccttcacca agtaccccgc cgacatccag
     4201 gactacttca agcagacctt ccccgagggc taccactggg agaggagcat gacctacgag
     4261 gaccagggca tctgcaccgc caccagcaac atcagcatga ggggcgactg cttcttctac
     4321 gacatcaggt tcgacggcac caacttcccc cccaacggcc ccgtgatgca gaagaagacc
     4381 ctgaagtggg agcccagcac cgagaagatg tacgtggagg acggcgtgct gaagggcgac
     4441 gtgaacatga ggctgctgct ggagggcggc ggccactaca ggtgcgactt caagaccacc
     4501 tacaaggcca agaaggaggt gaggctgccc gacgcccaca agatcgacca caggatcgag
     4561 atcctgaagc acgacaagga ctacaacaag gtgaagctgt acgagaacgc cgtggccagg
     4621 tactccatgc tgcccagcca ggccggatcc taactcgagg atccagacat gataagatac
     4681 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     4741 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaacg ctgagcatga
     4801 gacggaaatc tgctcgtcag tggtgctcac actgacgaat catgtacaga tcataccgat
     4861 gactgcctgg cgactcacaa ctaagcaaga cagccggaac cagcgccggc gaacaccact
     4921 gcatatatgg catatcacaa cagtccaact agtgcactgc agtaca
//
