LOCUS       sequence-418194-        3543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU2-b 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  JOURNAL   unpublished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            "unpublished"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: pTU2-b_UrK_amilCP
FEATURES             Location/Qualifiers
     source          1..3543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(122..916)
                     /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(233..252)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     843..862
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     terminator      complement(1055..1098)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1121..1155
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             1164..1175
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1848..1919
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(1942..2036)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2350..2541
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2535..2557)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2641..2780
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2696..2715
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2788..2805)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(2959..3543,1..4))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3039..3058)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtgaacga aaactcacgt
       61 taaagcaagc aaagaaaaaa ggcgacagat tactctgtcg ccttttttcc tgactcataa
      121 ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga
      181 taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagttcttca gcaatatcac
      241 gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga
      301 atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg ccgtgggtca
      361 cgacgagatc ctcgccgtcg ggcatacgcg ccttgagcct ggcgaacagt tcggctggcg
      421 cgagcccctg atgatcttcg tccagatcat cctgatcgac aagaccggct tccatccgag
      481 tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa
      541 gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt
      601 gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt
      661 cagtgacaac gtcgagcaca gccgcgcaag gaacgcccgt cgtggccagc cacgatagcc
      721 gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa
      781 ccgggcgacc ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt
      841 gtgcccagtc atagccgaat agcctctcca cccaagccgc cggagaacct gcgtgcaatc
      901 catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcagat cttgatcccc
      961 tgcgccatca gatccttggc ggcaagaaag ccatccagtt tactttgcag ggcttcccaa
     1021 ccttaccaga gggcgcccca gctggcaatt ccggatttca gataaaaaaa atccttagct
     1081 ttcgctaagg atgatttcgg tctcactata tctagagacg tttacagcta gctcagtcct
     1141 aggtattatg ctagctacta gagaaagagg agaaatacta gatgagtgtg atcgctaaac
     1201 aaatgaccta caaggtttat atgtcaggca cggtcaatgg acactacttt gaggtcgaag
     1261 gcgatggaaa aggtaagccc tacgaggggg agcagacggt aaagctcact gtcaccaagg
     1321 gcggacctct gccatttgct tgggatattt tatcaccaca gtgtcagtac ggaagcatac
     1381 cattcaccaa gtaccctgaa gatatccctg actatgtaaa gcagtcattc ccggagggct
     1441 atacatggga gaggatcatg aactttgaag atggtgcagt gtgtactgtc agcaatgatt
     1501 ccagcatcca aggcaactgt ttcatctacc atgtcaagtt ctctggtttg aactttcctc
     1561 ccaatggacc tgtcatgcag aagaaaacac agggctggga acccaacact gagcgtctgt
     1621 ttgcacgaga tggaatgctg ctaggaaaca actttatggc tctgaagtta gaaggaggcg
     1681 gtcactattt gtgtgaattt aaaactactt acaaggcaaa gaagcctgtg aagatgccag
     1741 ggtatcacta tgttgaccgc aaactggatg taaccaatca caacaaggat tacacttcgg
     1801 ttgagcagtg tgaaatttcc attgcacgca aacctgtggt cgcctaacaa ataaaacgaa
     1861 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     1921 gtctcagaag gtacagagac cattctcacc aataaaaaac gcccggcggc aaccgagcgt
     1981 tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca ggagtcctgc
     2041 ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag cgccctgcac cattatgttc
     2101 cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg
     2161 aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca taccgccagt
     2221 tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg
     2281 agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc cccttacacg
     2341 gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc
     2401 cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca ggcagacatc
     2461 tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt
     2521 gatgacggtg aaaacctctg acacatgcag ctcccggacg gtcacagctt gtctgtaagc
     2581 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     2641 cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca
     2701 tcagagcaga ttgtactgag agtgcaccat tgcggtgtga aataccgcac agatgcgtaa
     2761 ggagaaaata ccgcatcagg ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2821 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2881 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2941 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3001 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3061 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3121 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3181 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3241 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3301 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3361 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3421 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3481 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3541 atc
//
