LOCUS       sequence-451677-        2826 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU2-b backbone with kanamycin resistance and pUC ORI. amilCP 
            dropout vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2826)
  AUTHORS   Shapiro M
  TITLE     EcoFlex MoClo Extension, Volume I
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 2826)
  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: 
            "unpublished"
COMMENT     Sequence Label: pTU2-b_UK_amilCP
FEATURES             Location/Qualifiers
     source          1..2826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(118..912)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEDHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(229..248)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     839..858
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     terminator      complement(1051..1094)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1117..1151
                     /label=BBa_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     RBS             1160..1171
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1844..1915
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(1938..2032)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2067..2084)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2238..2826)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2318..2337)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt gaacgaaaac tcacgttaaa
       61 gcaagcaaag aaaaaaggcg acagattact ctgtcgcctt ttttcctgac tcataactca
      121 gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc
      181 gtaaagcacg aggaagcggt cagcccattc gccgccaagt tcttcagcaa tatcacgggt
      241 agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc
      301 agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccgt gggtcacgac
      361 gagatcctcg ccgtcgggca tacgcgcctt gagcctggcg aacagttcgg ctggcgcgag
      421 cccctgatga tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg
      481 tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt
      541 atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga
      601 tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt
      661 gacaacgtcg agcacagccg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc
      721 tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg
      781 gcgaccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc
      841 ccagtcatag ccgaatagcc tctccaccca agccgccgga gaacctgcgt gcaatccatc
      901 ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcagatcttg atcccctgcg
      961 ccatcagatc cttggcggca agaaagccat ccagtttact ttgcagggct tcccaacctt
     1021 accagagggc gccccagctg gcaattccgg atttcagata aaaaaaatcc ttagctttcg
     1081 ctaaggatga tttcggtctc actatatcta gagacgttta cggctagctc agccctaggt
     1141 attatgctag ctactagaga aagaggagaa atactagatg agtgtgatcg ctaaacaaat
     1201 gacctacaag gtttatatgt caggcacggt caatggacac tactttgagg tcgaaggcga
     1261 tggaaaaggt aagccctacg agggggagca gacggtaaag ctcactgtca ccaagggcgg
     1321 acctctgcca tttgcttggg atattttatc accacagtgt cagtacggaa gcataccatt
     1381 caccaagtac cctgaagata tccctgacta tgtaaagcag tcattcccgg agggctatac
     1441 atgggagagg atcatgaact ttgaagatgg tgcagtgtgt actgtcagca atgattccag
     1501 catccaaggc aactgtttca tctaccatgt caagttctct ggtttgaact ttcctcccaa
     1561 tggacctgtc atgcagaaga aaacacaggg ctgggaaccc aacactgagc gtctgtttgc
     1621 acgagatgga atgctgctag gaaacaactt tatggctctg aagttagaag gaggcggtca
     1681 ctatttgtgt gaatttaaaa ctacttacaa ggcaaagaag cctgtgaaga tgccagggta
     1741 tcactatgtt gaccgcaaac tggatgtaac caatcacaac aaggattaca cttcggttga
     1801 gcagtgtgaa atttccattg cacgcaaacc tgtggtcgcc taacaaataa aacgaaaggc
     1861 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctccgtct
     1921 cagaaggtac agagaccatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg
     1981 aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag tcctgcggtt
     2041 tccgtgtttc gtaaagtctg ggcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2101 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2161 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2221 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2281 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2341 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2401 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2461 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2521 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2581 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2641 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2701 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2761 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2821 tctcaa
//
