LOCUS       sequence-206339-        2639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Aquamarine (an eCFP derivative) in Ecoli off a T7 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2639)
  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 2639)
  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: pJL1 Aquamarine
FEATURES             Location/Qualifiers
     source          1..2639
                     /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"
     primer_bind     617..636
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        617..635
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             667..689
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             709..726
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             748..768
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(772..774,775..1488)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISGNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(814..835)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1075..1094)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1422..1443
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1624..1642)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1638..1685
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1703..2518)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2426..2445
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2510..2529
                     /label=pENTR-R
                     /note="pENTR vectors, 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 cgcgatcccg cgaaattaat acgactcact atagggagac cacaacggtt tccctctaga
      661 aataattttg tttaacttta agaaggagat atacatatgt cgtactacca tcaccatcac
      721 catcacgatt acgatatccc aacgaccgaa aacctgtatt ttcagggcgc cgtgagcaag
      781 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      841 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      901 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      961 ctgagctggg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     1021 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     1081 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1141 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1201 aactacatca gcggcaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc
     1261 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1321 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1381 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1441 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag gatcccggga
     1501 attctcgagt aaggttaacc tgcaggaggc ctttaattaa ggtggtgcgg ccgcgctagc
     1561 ggtcccgggg gatcgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     1621 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     1681 ttttgctgaa agccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt
     1741 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     1801 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     1861 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     1921 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta tgcatttctt
     1981 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     2041 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     2101 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     2161 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     2221 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     2281 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     2341 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     2401 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     2461 ccatgttgga atttaatcgc ggcttcgagc aagacgtttc ccgttgaata tggctcataa
     2521 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt
     2581 tatcttgtgc aatgtaacat cagagatttt gagacacaac gtgagatcaa aggatcttc
//
