LOCUS       sequence-252478-        4334 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes TU with connectors L1 R2 and an B1 insulator, and pCMV 
            expressing cytoplasmic mAzamiGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4334)
  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 4334)
  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: ARB346
FEATURES             Location/Qualifiers
     source          1..4334
                     /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        2325..2628
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2629..2832
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2783..2803
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2880..2906
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             2913..3584
                     /codon_start=1
                     /product="humanized monomeric Azami-Green fluorescent 
                     protein"
                     /label=hmAzami-Green
                     /note="mammalian codon-optimized"
                     /translation="VSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNVT
                     EGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTAT
                     SNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLLL
                     EGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLPS
                     QA"
     CDS             2913..3584
                     /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(3681..3700)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3746..3801
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3800..3819)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
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 ctagaacggg gtcatcacgg ctcatcatgc gccaaacaaa
     1921 tgtgtgcaat acacgctcgg atgactgcat gatgaccgca ctgactgggg acagcagatc
     1981 cacctaagcc tgtgagagaa gcagacaccc gacagatcaa ggcagttacg tctcaccaaa
     2041 ccagatgtca acacagctac ggtctgccgc ttcaggtcca tttacggaag ccgcaatcct
     2101 gcactatgac caccaggggg agcgctaggc ccatccctgc acaggtagct atttttagag
     2161 gttgcttcta gcctggagga agaaacgaaa gaaataaaag actatgatac tttttttttt
     2221 taaccggacc aacttataaa ttatttgtaa gtccaataaa aacgtagtta ttaatagtaa
     2281 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     2341 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     2401 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     2461 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     2521 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     2581 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     2641 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     2701 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     2761 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     2821 ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc gccactatgc
     2881 tgccccccct ggagaggctg accctgggtt ctgtgagcgt gatcaagccc gagatgaaga
     2941 tcaagctgtg catgaggggc accgtgaacg gccacaactt cgtgatcgag ggcgagggca
     3001 agggcaaccc ctacgagggc acccagatcc tggacctgaa cgtgaccgag ggcgcccccc
     3061 tgcccttcgc ctacgacatc ctgaccaccg tgttccagta cggcaacagg gccttcacca
     3121 agtaccccgc cgacatccag gactacttca agcagacctt ccccgagggc taccactggg
     3181 agaggagcat gacctacgag gaccagggca tctgcaccgc caccagcaac atcagcatga
     3241 ggggcgactg cttcttctac gacatcaggt tcgacggcac caacttcccc cccaacggcc
     3301 ccgtgatgca gaagaagacc ctgaagtggg agcccagcac cgagaagatg tacgtggagg
     3361 acggcgtgct gaagggcgac gtgaacatga ggctgctgct ggagggcggc ggccactaca
     3421 ggtgcgactt caagaccacc tacaaggcca agaaggaggt gaggctgccc gacgcccaca
     3481 agatcgacca caggatcgag atcctgaagc acgacaagga ctacaacaag gtgaagctgt
     3541 acgagaacgc cgtggccagg tactccatgc tgcccagcca ggccggatcc taactcgaga
     3601 ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg
     3661 gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag
     3721 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     3781 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
     3841 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
     3901 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
     3961 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
     4021 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
     4081 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag
     4141 cccaagcttg gcgtaatgct ggatgtgaga cgataaatcg cagccaagtg agtgaataga
     4201 tgacgcacca cggtcagaca cggccacatc gtatctcaca ggagcaagcg cgataggagc
     4261 actcacacat agtacggtga tccgctgact cctttgccca aataagacgt gagccactag
     4321 tgcactgcag taca
//
