LOCUS       sequence-204385-        3000 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Aquamarine (an eCFP derivative) in Ecoli off a T7 promoter
            when pNP1 Aqua trigger is co-expressed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3000)
  AUTHORS   Huang A, Nguyen PQ, Stark JC, Takahashi MK, Donghia N, Ferrante T, 
            Dy AJ, Hsu KJ, Dubner RS, Pardee K, Jewett MC, Collins JJ
  TITLE     BioBits Explorer: A modular synthetic biology education kit.
  JOURNAL   Sci Adv. 2018 Aug 1;4(8):eaat5105. doi: 10.1126/sciadv.aat5105. 
            eCollection 2018 Aug.
  PUBMED    30083608
REFERENCE   2  (bases 1 to 3000)
  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: "Sci Adv."; 
            date: "2018-08-1"; volume: "4(8):eaat5105. doi"; pages: " 
            10.1126/sciadv.aat5105. eCollection 2018 Aug"
COMMENT     Sequence Label: pNP1 Aqua toehold switch
FEATURES             Location/Qualifiers
     source          1..3000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     26..45
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        26..44
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(126..128,129..842)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISGNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(168..189)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(429..448)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     776..797
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(978..996)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      992..1039
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(1257..1845)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1337..1356)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2017..2808,2809..2877))
                     /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     2640..2659
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2878..2982)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gctagtggtg ctagccccgc gaaattaata cgactcacta tagggacaaa gattggtcgt
       61 ttcattaccg ttagaaacag aggagatcta acatgaatga aacgaacctg gcggcagcgc
      121 aaaaggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
      181 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      241 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
      301 ccctcgtgac caccctgagc tggggcgtgc agtgcttcag ccgctacccc gaccacatga
      361 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
      421 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
      481 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
      541 acaagctgga gtacaactac atcagcggca acgtctatat caccgccgac aagcagaaga
      601 acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
      661 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
      721 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
      781 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
      841 aaaggatccc gggaattctc gagtaaggtt aacctgcagg aggcctttaa ttaaggtggt
      901 gcggccgcgc tagcggtccc gggggatcga tccggctgct aacaaagccc gaaaggaagc
      961 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     1021 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggaagcttgg cactggccga
     1081 ccggggtcga gcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     1141 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1201 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1261 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1321 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1381 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1441 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     1501 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     1561 cgtcttgagt ccaacccgct aagacacgac ttatcgccac tggcagcagc cactggtaac
     1621 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1681 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1741 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1801 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1861 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac agatccggga ttttggtcat
     1921 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     1981 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2041 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2101 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2161 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2221 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2281 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2341 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2401 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     2461 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     2521 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     2581 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     2641 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     2701 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     2761 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     2821 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     2881 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     2941 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
//
