LOCUS       sequence-114408-        3092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Transcriptional Unit: FACS Standard Color Controls - High GFP 
            expression cassette - Ampicillin plasmid 
            [E:J23102:B:BCD2:C:E0040m:D:B0015:F].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3092)
  AUTHORS   Iverson SV, Haddock TL, Beal J, Densmore DM
  TITLE     CIDAR MoClo: Improved MoClo Assembly Standard and New E. coli Part 
            Library Enable Rapid Combinatorial Design for Synthetic and 
            Traditional Biology.
  JOURNAL   ACS Synth Biol. 2016 Jan 15;5(1):99-103. doi: 
            10.1021/acssynbio.5b00124. Epub 2015 Nov 4.
  PUBMED    26479688
REFERENCE   2  (bases 1 to 3092)
  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.5b00124"; journalName: "ACS Synth Biol"; date: 
            "2016-01-15- 15"; volume: "5"; issue: "1"; pages: "99-103"
COMMENT     Sequence Label: pJ02B2Gm_EF
FEATURES             Location/Qualifiers
     source          1..3092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..39
                     /label=BBa_J23102
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.86"
     RBS             114..122
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             132..845
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(162..190)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     785..805
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      868..939
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      955..982
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1000..1020
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(1026..1043)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1197..1785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1277..1296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1956..2747,2748..2816))
                     /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     2579..2598
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2817..2921)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2989..3007
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3011..3054)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3057..3078
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 gcttttgaca gctagctcag tcctaggtac tgtgctagct actgggccca agttcactta
       61 aaaaggagat caacaatgaa agcaattttc gtactgaaac atcttaatca tgctaaggag
      121 gttttctaat gatgcgtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg
      181 aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg
      241 caacatacgg aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat
      301 ggccaacact tgtcactact ttcggttatg gtgttcaatg ctttgcgaga tacccagatc
      361 atatgaaaca gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa
      421 ctatattttt caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg
      481 atacccttgt taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc
      541 ttggacacaa attggaatac aactataact cacacaatgt atacatcatg gcagacaaac
      601 aaaagaatgg aatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggaagcgttc
      661 aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag
      721 acaaccatta cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagagagatc
      781 acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat
      841 acaaataata aaggtaggtc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
      901 gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca
      961 ccttcgggtg ggcctttctg cgtttatacg ctagagacct actagtagcg gccgctgcag
     1021 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     1081 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1141 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1201 cacaggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1261 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1321 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1381 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     1441 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     1501 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     1561 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1621 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1681 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1741 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1801 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1861 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1921 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1981 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     2041 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgcgac
     2101 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     2161 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     2221 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     2281 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     2341 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     2401 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     2461 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     2521 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     2581 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     2641 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     2701 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     2761 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     2821 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     2881 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     2941 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     3001 acgaggcaga atttcagata aaaaaaatcc ttagctttcg ctaaggatga tttctggaat
     3061 tcgcggccgc ttctagagac tagtgggtct ca
//
