LOCUS       sequence-418175-        4850 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU2-a backbone with kanamycin resistance, pSC101-var2 ORI, and 
            Axe-Txe plasmid stability cassette. amilCP dropout vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1557 to 1591)
  AUTHORS   Shapiro M
  TITLE     EcoFlex MoClo Extension, Volume I
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 4850)
  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            "unpublished"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-a_Sv2KA_amilCP
FEATURES             Location/Qualifiers
     source          1..4850
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(523..1317)
                     /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(634..653)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1244..1263
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     terminator      complement(1491..1534)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1557..1591
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     RBS             1600..1611
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2284..2356
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3595..4545)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFWIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      4593..4815
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 tgtcgcaaac gctgtttgct cctctacaaa acagacctta aaaccctaaa ggcttaagta
       61 gcaccctcgc aagctcggtt gcggccgcaa tcgggcaaat cgctgaatat tccttttgtc
      121 tccgaccatc aggcacctga gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac
      181 ggctctggca gtgaatgggg gtaaatggca ctacaggcgc cttttatgga ttcatgcaag
      241 gaaactaccc ataatacaag aaaagcccgt cacgggcttc caggcgttta ttggcgggtc
      301 tgctatgtgg tgctatctga ctttttgctg ttcagcagtt cctgccctct gattttccag
      361 tctgaccact tcggattatc ccgtgacagg tcattcagac tggctaatgc acccagtaag
      421 gcagcggtat catcaacaag caaagaaaaa aggcgacaga ttactctgtc gccttttttc
      481 ctgactcata accaagccaa ttctcgaacc ccagagtccc gctcagaaga actcgtcaag
      541 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
      601 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
      661 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
      721 ttccaccatg atattcggca agcaggcatc gccgtgggtc acgacgagat cctcgccgtc
      781 gggcatacgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgatcttc
      841 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
      901 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
      961 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     1021 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     1081 agccgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag
     1141 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgac cctgcgctga
     1201 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     1261 tagcctctcc acccaagccg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg
     1321 aaacgatcct catcctgtct cttgatcaga tcttgatccc ctgcgccatc agatccttgg
     1381 cggcaagaaa gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc
     1441 agctggcaat tccggcaccc cttgtattac tgtttatgta agcagacagt atttcagata
     1501 aaaaaaatcc ttagctttcg ctaaggatga tttcggtctc actatatcta gagacgttga
     1561 cggctagctc agtcctaggt acagtgctag ctactagaga aagaggagaa atactagatg
     1621 agtgtgatcg ctaaacaaat gacctacaag gtttatatgt caggcacggt caatggacac
     1681 tactttgagg tcgaaggcga tggaaaaggt aagccctacg agggggagca gacggtaaag
     1741 ctcactgtca ccaagggcgg acctctgcca tttgcttggg atattttatc accacagtgt
     1801 cagtacggaa gcataccatt caccaagtac cctgaagata tccctgacta tgtaaagcag
     1861 tcattcccgg agggctatac atgggagagg atcatgaact ttgaagatgg tgcagtgtgt
     1921 actgtcagca atgattccag catccaaggc aactgtttca tctaccatgt caagttctct
     1981 ggtttgaact ttcctcccaa tggacctgtc atgcagaaga aaacacaggg ctgggaaccc
     2041 aacactgagc gtctgtttgc acgagatgga atgctgctag gaaacaactt tatggctctg
     2101 aagttagaag gaggcggtca ctatttgtgt gaatttaaaa ctacttacaa ggcaaagaag
     2161 cctgtgaaga tgccagggta tcactatgtt gaccgcaaac tggatgtaac caatcacaac
     2221 aaggattaca cttcggttga gcagtgtgaa atttccattg cacgcaaacc tgtggtcgcc
     2281 taacaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgtttgttt
     2341 gtcggtgaac gctctccgtc tcaccgggta cagagaccaa gcgtcatcct tagtaactaa
     2401 catcccacac tacaatagta ttataacata tctattcaac ttcaaattgt tgaaatttaa
     2461 gccatttata tttacttttc ctcaacttta gaaatgattt tactaacctt atatccgtaa
     2521 tacgcgtaac aaacacattc attaataaca atcaacacta agctatatta gctttttaaa
     2581 tgactagggt aaataaaata aggatcaaaa acttttaagt ttctgaccct ttccttactt
     2641 ccgattggtt aatagtgatc ttttgcagaa taaataaata tcgtttcatt ttcaactcta
     2701 tatatcagtc tatgttcatc tgtaattctt ctggaccatt ttccagataa atcatgcttt
     2761 aatggctcag gttttcctaa tccagcaaag ggggaacgat cgatatcttt tattaacttg
     2821 ttaatctttt ttatattgct tttgtttcct tgctcatgcc aataaagata atcatcccaa
     2881 gcatcatcag accaagcctt aatcatcaga ttcaacctcg attaagtcat gtgttttaaa
     2941 tgcacctttg gagaattgtt catctcctcg acgaattttt tccatgacgt aattattaga
     3001 aagtgttctc aacgtttctt gcatagaatc ataatctctt tttgataata caacaactgt
     3061 atcttctaca tctttacttg ttacaataag tgtttcagca tcctcattaa cttgtttcat
     3121 ataactacgt aaattttggc ggaaatttga ataagctact gcttccattc ctttcacccc
     3181 ttatttattt ctataaaaca attgtacatt atattgtaca attaagcaaa tgattaatat
     3241 tctaacttct tataattaaa agtgctatca cttttagttt ttaggataaa aagtgacgtc
     3301 actttttgag tgctggtacc tcaaatcctg ctttttcttg ggcaatcaat aaatctaacg
     3361 caatacacat aatcctggat tcacttttgc cgtacttctg ctttagttgc tgcaattctt
     3421 tttgtctctt ttcatccaac gtaacgggta accgtttatt tcctattgca attggcatta
     3481 gtactctcct agtctatgga cagttttccc tttgatatgt aacggtgaac agttgttcta
     3541 cttttgtttg ttagtcttga tgcttcactg atagatacaa gagccataag aacctcagat
     3601 ccttccgtat ttagccagta tgttctctag tgtggttcgt tgtttttgcg tgagccatga
     3661 gaacgaacca ttgagatcat acttactttg catgtcactc aaaaattttg cctcaaaact
     3721 ggtgagctga atttttgcag ttaaagcatc gtgtagtgtt tttcttagtc cgttatgtag
     3781 gtaggaatct gatgtaatgg ttgttggtat tttgtcacca ttcattttta tctggttgtt
     3841 ctcaagttcg gttacgagat ccatttgtct atctagttca acttggaaaa tcaacgtatc
     3901 agtcgggcgg cctcgcttat caaccaccaa tttcatattg ctgtaagtgt ttaaatcttt
     3961 acttattggt ttcaaaaccc attggttaag ccttttaaac tcatggtagt tattttcaag
     4021 cattaacatg aacttaaatt catcaaggct aatctctata tttgccttgt gagttttctt
     4081 ttgtgttagt tcttttaata accactcata aatcctcata gagtatttgt tttcaaaaga
     4141 cttaacatgt tccagattat attttatgaa tttttttaac tggaaaagat aaggcaatat
     4201 ctcttcacta aaaactaatt ctaatttttc gcttgagaac ttggcatagt ttgtccactg
     4261 gaaaatccaa aagcctttaa ccaaaggatt cctgatttcc acagttctcg tcatcagctc
     4321 tctggttgct ttagctaata caccataagc attttcccta ctgatgttca tcatctgagc
     4381 gtattggtta taagtgaacg ataccgtccg ttctttcctt gtagggtttt caatcgtggg
     4441 gttgagtagt gccacacagc ataaaattag cttggtttca tgctccgtta agtcatagcg
     4501 actaatcgct agttcatttg ctttgaaaac aactaattca gacatacatc tcaattggtc
     4561 taggtgattt taatcactat accaattgag atgggctagt caatgataat tactagtcct
     4621 tttcctttga gttgtgggta tctgtaaatt ctgctagacc tttgctggaa aacttgtaaa
     4681 ttctgctaga ccctctgtaa attccgctag acctttgtgt gttttttttg tttatattca
     4741 agtggttata atttatagaa taaagaaaga ataaaaaaag ataaaaagaa tagatcccag
     4801 ccctgtgtat aactcactac tttagtcagt tccgcagtat tacaaaagga
//
