LOCUS       sequence-231160-        3078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo golden gate assembly AE destination vector DVCK-AE-GFP 
            (ColE1/kan resistance destination vector with constitutive GFP 
            expression for screening..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3078)
  TITLE     CIDAR MoClo Extension
REFERENCE   2  (bases 1 to 3078)
  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"
COMMENT     Sequence Label: V35m
FEATURES             Location/Qualifiers
     source          1..3078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..24
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(28..71)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    74..95
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        159..193
                     /label=BBa_J23102
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.86"
     CDS             224..937
                     /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(254..282)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     877..897
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      956..1027
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1043..1070
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1180..1200
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1201..1258
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(1282..1299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1507..1526)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1518..2333
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1591..1610)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(2455..3043)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2535..2554)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcataaatag gcgtatcacg aggcagaatt tcagataaaa aaaatcctta gctttcgcta
       61 aggatgattt ctggaattcg cggccgcttc tagagactag tggaagacat ggagagagac
      121 ctgcaccata tgcggtgaga ctagtggaag acatggagtt gacagctagc tcagtcctag
      181 gtactgtgct agctactaga gtcacacagg aaagtactaa atgatgcgta aaggagaaga
      241 acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta atgggcacaa
      301 attttctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta cccttaaatt
      361 tatttgcact actggaaaac tacctgttcc atggccaaca cttgtcacta ctttcggtta
      421 tggtgttcaa tgctttgcga gatacccaga tcatatgaaa cagcatgact ttttcaagag
      481 tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg acgggaacta
      541 caagacacgt gctgaagtca agtttgaagg tgataccctt gttaatagaa tcgagttaaa
      601 aggtattgat tttaaagaag atggaaacat tcttggacac aaattggaat acaactataa
      661 ctcacacaat gtatacatca tggcagacaa acaaaagaat ggaatcaaag ttaacttcaa
      721 aattagacac aacattgaag atggaagcgt tcaactagca gaccattatc aacaaaatac
      781 tccaattggc gatggccctg tccttttacc agacaaccat tacctgtcca cacaatctgc
      841 cctttcgaaa gatcccaacg aaaagagaga tcacatggtc cttcttgagt ttgtaacagc
      901 tgctgggatt acacatggca tggatgaact atacaaataa taaaggtcca ggcatcaaat
      961 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     1021 cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttatagctt
     1081 atgtcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     1141 tgaatcggcc aacgcgcggg ggtctctgct tatgtcttct actagtagcg gccgctgcag
     1201 tccggcaaaa aagggcaagg tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt
     1261 acttcgcgtt atgcaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     1321 gcgagcggta tcagctcact caaaggcggt aagtcgcgtg ttatacgccc gttatccatg
     1381 ggtatggaca gttttccctt tgatatgtaa cgcacgttgt gtctcaaaat ctctgatgtt
     1441 acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca
     1501 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcttgctcc cgtccgcgct
     1561 taaactccaa catggacgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     1621 aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga
     1681 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtccgt ctcaactggc
     1741 tgacggagtt tatgcctctc ccgaccatca agcattttat ccgtactcct gatgatgcgt
     1801 ggttactcac caccgcgatt cctgggaaaa cagccttcca ggtattagaa gaatatcctg
     1861 attcaggtga aaatattgtt gatgcgctgg ccgtgttcct gcgccggtta cattcgattc
     1921 ctgtttgtaa ttgtcctttt aacagcgatc gtgtatttcg tcttgctcag gcgcaatcac
     1981 gcatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     2041 ttgaacaagt ctggaaagaa atgcacaagc tcttgccatt ctcaccggat tcagtcgtca
     2101 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     2161 ttgatgttgg acgggtcgga atcgcagacc gttaccagga ccttgccatt ctttggaact
     2221 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     2281 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taataatact
     2341 agcagaaatc atccttagcg aaagctaagg atttttttta tctgattacc gcctttgagt
     2401 gagcgtcgac ctagtcctag ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2461 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2521 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2581 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2641 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2701 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2761 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2821 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2881 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2941 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3001 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3061 ttctacgggg tctgacgc
//
