LOCUS       sequence-418162-        3543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU1-A backbone with kanamycin resistance and pUC-bom-rop ORI. 
            amilCP dropout vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3543)
  AUTHORS   Shapiro M
  TITLE     EcoFlex MoClo Extension, Volume I
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 3543)
  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            "unpublished"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: pTU1-A_UrK_amilCP
FEATURES             Location/Qualifiers
     source          1..3543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(119..913)
                     /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(230..249)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     840..859
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     terminator      complement(1052..1095)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1118..1152
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             1161..1172
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1845..1916
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(1939..2033)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2347..2538
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2532..2554)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2638..2777
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2693..2712
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2785..2802)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(2956..3543,1))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3036..3055)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agaagatcct ttgatctttt ctacggggtc tgacgctcag tgaacgaaaa ctcacgttaa
       61 agcaagcaaa gaaaaaaggc gacagattac tctgtcgcct tttttcctga ctcataactc
      121 agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac
      181 cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ttcttcagca atatcacggg
      241 tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc
      301 cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgccg tgggtcacga
      361 cgagatcctc gccgtcgggc atacgcgcct tgagcctggc gaacagttcg gctggcgcga
      421 gcccctgatg atcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac
      481 gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg
      541 tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag
      601 atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag
      661 tgacaacgtc gagcacagcc gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg
      721 ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg
      781 ggcgaccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg
      841 cccagtcata gccgaatagc ctctccaccc aagccgccgg agaacctgcg tgcaatccat
      901 cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatctt gatcccctgc
      961 gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc ttcccaacct
     1021 taccagaggg cgccccagct ggcaattccg gatttcagat aaaaaaaatc cttagctttc
     1081 gctaaggatg atttccgtct caatctctat agagaccttt acagctagct cagtcctagg
     1141 tattatgcta gctactagag aaagaggaga aatactagat gagtgtgatc gctaaacaaa
     1201 tgacctacaa ggtttatatg tcaggcacgg tcaatggaca ctactttgag gtcgaaggcg
     1261 atggaaaagg taagccctac gagggggagc agacggtaaa gctcactgtc accaagggcg
     1321 gacctctgcc atttgcttgg gatattttat caccacagtg tcagtacgga agcataccat
     1381 tcaccaagta ccctgaagat atccctgact atgtaaagca gtcattcccg gagggctata
     1441 catgggagag gatcatgaac tttgaagatg gtgcagtgtg tactgtcagc aatgattcca
     1501 gcatccaagg caactgtttc atctaccatg tcaagttctc tggtttgaac tttcctccca
     1561 atggacctgt catgcagaag aaaacacagg gctgggaacc caacactgag cgtctgtttg
     1621 cacgagatgg aatgctgcta ggaaacaact ttatggctct gaagttagaa ggaggcggtc
     1681 actatttgtg tgaatttaaa actacttaca aggcaaagaa gcctgtgaag atgccagggt
     1741 atcactatgt tgaccgcaaa ctggatgtaa ccaatcacaa caaggattac acttcggttg
     1801 agcagtgtga aatttccatt gcacgcaaac ctgtggtcgc ctaacaaata aaacgaaagg
     1861 atcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcggtc
     1921 tcatgtttgc cagagacgat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct
     1981 gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga gtcctgcggt
     2041 ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg
     2101 atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt attaacgaag
     2161 cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac cgccagttgt
     2221 ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg tatcgtgagc
     2281 atcctctctc gtttcatcgg tatcattacc cccatgaaca gaaatccccc ttacacggag
     2341 gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat cagaagccag
     2401 acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc agacatctgt
     2461 gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg tttcggtgat
     2521 gacggtgaaa acctctgaca catgcagctc ccggacggtc acagcttgtc tgtaagcgga
     2581 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     2641 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
     2701 gagcagattg tactgagagt gcaccattgc ggtgtgaaat accgcacaga tgcgtaagga
     2761 gaaaataccg catcaggccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2821 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2881 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2941 cgcgttgctg gcgtttttcc acaggctccg cccccctgac gagcatcaca aaaatcgacg
     3001 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3061 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3121 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3181 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3241 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3301 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3361 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3421 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3481 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3541 tca
//
