LOCUS       sequence-418172-        4851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU2-a backbone with kanamycin resistance, pSC101-var3 ORI, and 
            Axe-Txe plasmid stability cassette. amilCP dropout vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1554 to 1588)
  AUTHORS   Shapiro M
  TITLE     EcoFlex MoClo Extension, Volume I
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 4851)
  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            "unpublished"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_Sv3KA_amilCP
FEATURES             Location/Qualifiers
     source          1..4851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(520..1314)
                     /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(631..650)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1241..1260
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     terminator      complement(1488..1531)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1554..1588
                     /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             1597..1608
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2281..2352
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3591..4541)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFKIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      4589..4811
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 caaacgctgt ttgctcctct acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc
       61 ctcgcaagct cggttgcggc cgcaatcggg caaatcgctg aatattcctt ttgtctccga
      121 ccatcaggca cctgagtcgc tgtctttttc gtgacattca gttcgctgcg ctcacggctc
      181 tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt atggattcat gcaaggaaac
      241 tacccataat acaagaaaag cccgtcacgg gcttctcagg gcgttttatg gcgggtctgc
      301 tatgtggtgc tatctgactt tttgctgttc agcagttcct gccctctgat tttccagtct
      361 gaccacttcg gattatcccg tgacaggtca ttcagactgg ctaatgcacc cagtaaggca
      421 gcggtatcat caacaagcaa agaaaaaagg cgacagatta ctctgtcgcc ttttttcctg
      481 actcataacc aagccaattc tcgaacccca gagtcccgct cagaagaact cgtcaagaag
      541 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg
      601 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg
      661 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc
      721 caccatgata ttcggcaagc aggcatcgcc gtgggtcacg acgagatcct cgccgtcggg
      781 catacgcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gatcttcgtc
      841 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg
      901 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc
      961 atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc
     1021 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc
     1081 cgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cttgcagttc
     1141 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgaccct gcgctgacag
     1201 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag
     1261 cctctccacc caagccgccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa
     1321 cgatcctcat cctgtctctt gatcagatct tgatcccctg cgccatcaga tccttggcgg
     1381 caagaaagcc atccagttta ctttgcaggg cttcccaacc ttaccagagg gcgccccagc
     1441 tggcaattcc ggcacccctt gtattactgt ttatgtaagc agacagtatt tcagataaaa
     1501 aaaatcctta gctttcgcta aggatgattt cggtctcact atatctagag acgttgacgg
     1561 ctagctcagt cctaggtaca gtgctagcta ctagagaaag aggagaaata ctagatgagt
     1621 gtgatcgcta aacaaatgac ctacaaggtt tatatgtcag gcacggtcaa tggacactac
     1681 tttgaggtcg aaggcgatgg aaaaggtaag ccctacgagg gggagcagac ggtaaagctc
     1741 actgtcacca agggcggacc tctgccattt gcttgggata ttttatcacc acagtgtcag
     1801 tacggaagca taccattcac caagtaccct gaagatatcc ctgactatgt aaagcagtca
     1861 ttcccggagg gctatacatg ggagaggatc atgaactttg aagatggtgc agtgtgtact
     1921 gtcagcaatg attccagcat ccaaggcaac tgtttcatct accatgtcaa gttctctggt
     1981 ttgaactttc ctcccaatgg acctgtcatg cagaagaaaa cacagggctg ggaacccaac
     2041 actgagcgtc tgtttgcacg agatggaatg ctgctaggaa acaactttat ggctctgaag
     2101 ttagaaggag gcggtcacta tttgtgtgaa tttaaaacta cttacaaggc aaagaagcct
     2161 gtgaagatgc cagggtatca ctatgttgac cgcaaactgg atgtaaccaa tcacaacaag
     2221 gattacactt cggttgagca gtgtgaaatt tccattgcac gcaaacctgt ggtcgcctaa
     2281 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     2341 gtgaacgctc tccgtctcac cgggtacaga gaccaagcgt catccttagt aactaacatc
     2401 ccacactaca atagtattat aacatatcta ttcaacttca aattgttgaa atttaagcca
     2461 tttatattta cttttcctca actttagaaa tgattttact aaccttatat ccgtaatacg
     2521 cgtaacaaac acattcatta ataacaatca acactaagct atattagctt tttaaatgac
     2581 tagggtaaat aaaataagga tcaaaaactt ttaagtttct gaccctttcc ttacttccga
     2641 ttggttaata gtgatctttt gcagaataaa taaatatcgt ttcattttca actctatata
     2701 tcagtctatg ttcatctgta attcttctgg accattttcc agataaatca tgctttaatg
     2761 gctcaggttt tcctaatcca gcaaaggggg aacgatcgat atcttttatt aacttgttaa
     2821 tcttttttat attgcttttg tttccttgct catgccaata aagataatca tcccaagcat
     2881 catcagacca agccttaatc atcagattca acctcgatta agtcatgtgt tttaaatgca
     2941 cctttggaga attgttcatc tcctcgacga attttttcca tgacgtaatt attagaaagt
     3001 gttctcaacg tttcttgcat agaatcataa tctctttttg ataatacaac aactgtatct
     3061 tctacatctt tacttgttac aataagtgtt tcagcatcct cattaacttg tttcatataa
     3121 ctacgtaaat tttggcggaa atttgaataa gctactgctt ccattccttt caccccttat
     3181 ttatttctat aaaacaattg tacattatat tgtacaatta agcaaatgat taatattcta
     3241 acttcttata attaaaagtg ctatcacttt tagtttttag gataaaaagt gacgtcactt
     3301 tttgagtgct ggtacctcaa atcctgcttt ttcttgggca atcaataaat ctaacgcaat
     3361 acacataatc ctggattcac ttttgccgta cttctgcttt agttgctgca attctttttg
     3421 tctcttttca tccaacgtaa cgggtaaccg tttatttcct attgcaattg gcattagtac
     3481 tctcctagtc tatggacagt tttccctttg atatgtaacg gtgaacagtt gttctacttt
     3541 tgtttgttag tcttgatgct tcactgatag atacaagagc cataagaacc tcagatcctt
     3601 ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac
     3661 gaaccattga gatcatactt actttgcatg tcactcaaaa attttgcctc aaaactggtg
     3721 agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt atgtaggtag
     3781 gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg gttgttctca
     3841 agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc
     3901 gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa atctttactt
     3961 attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt ttcaagcatt
     4021 aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt tttcttttgt
     4081 gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc aaaagactta
     4141 acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg caatatctct
     4201 tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt ccactggaaa
     4261 atcttaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat cagctctctg
     4321 gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat ctgagcgtat
     4381 tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat cgtggggttg
     4441 agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc atagcgacta
     4501 atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa ttggtctagg
     4561 tgattttaat cactatacca attgagatgg gctagtcaat gataattact agtccttttc
     4621 ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact tgtaaattct
     4681 gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta tattcaagtg
     4741 gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga tcccagccct
     4801 gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc g
//
