LOCUS       sequence-413881-        4950 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU2-a backbone with kanamycin resistance, pSC101 (WT) ORI, and 
            Axe-Txe plasmid stability cassette. amilCP dropout vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1634 to 1668)
  AUTHORS   Shapiro M
  TITLE     EcoFlex MoClo Extension, Volume I
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 4950)
  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            "unpublished"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_SKA_amilCP
FEATURES             Location/Qualifiers
     source          1..4950
                     /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(1568..1611)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1634..1668
                     /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             1677..1688
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2361..2432
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3690..4640)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      4688..4910
                     /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 agacagtttt attgttgcca
     1501 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     1561 agacagtatt tcagataaaa aaaatcctta gctttcgcta aggatgattt cggtctcact
     1621 atatctagag acgttgacgg ctagctcagt cctaggtaca gtgctagcta ctagagaaag
     1681 aggagaaata ctagatgagt gtgatcgcta aacaaatgac ctacaaggtt tatatgtcag
     1741 gcacggtcaa tggacactac tttgaggtcg aaggcgatgg aaaaggtaag ccctacgagg
     1801 gggagcagac ggtaaagctc actgtcacca agggcggacc tctgccattt gcttgggata
     1861 ttttatcacc acagtgtcag tacggaagca taccattcac caagtaccct gaagatatcc
     1921 ctgactatgt aaagcagtca ttcccggagg gctatacatg ggagaggatc atgaactttg
     1981 aagatggtgc agtgtgtact gtcagcaatg attccagcat ccaaggcaac tgtttcatct
     2041 accatgtcaa gttctctggt ttgaactttc ctcccaatgg acctgtcatg cagaagaaaa
     2101 cacagggctg ggaacccaac actgagcgtc tgtttgcacg agatggaatg ctgctaggaa
     2161 acaactttat ggctctgaag ttagaaggag gcggtcacta tttgtgtgaa tttaaaacta
     2221 cttacaaggc aaagaagcct gtgaagatgc cagggtatca ctatgttgac cgcaaactgg
     2281 atgtaaccaa tcacaacaag gattacactt cggttgagca gtgtgaaatt tccattgcac
     2341 gcaaacctgt ggtcgcctaa caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     2401 gttttatctg ttgtttgtcg gtgaacgctc tccgtctcac cgggtacaga gaccaagcgt
     2461 catccttagt aactaacatc ccacactaca atagtattat aacatatcta ttcaacttca
     2521 aattgttgaa atttaagcca tttatattta cttttcctca actttagaaa tgattttact
     2581 aaccttatat ccgtaatacg cgtaacaaac acattcatta ataacaatca acactaagct
     2641 atattagctt tttaaatgac tagggtaaat aaaataagga tcaaaaactt ttaagtttct
     2701 gaccctttcc ttacttccga ttggttaata gtgatctttt gcagaataaa taaatatcgt
     2761 ttcattttca actctatata tcagtctatg ttcatctgta attcttctgg accattttcc
     2821 agataaatca tgctttaatg gctcaggttt tcctaatcca gcaaaggggg aacgatcgat
     2881 atcttttatt aacttgttaa tcttttttat attgcttttg tttccttgct catgccaata
     2941 aagataatca tcccaagcat catcagacca agccttaatc atcagattca acctcgatta
     3001 agtcatgtgt tttaaatgca cctttggaga attgttcatc tcctcgacga attttttcca
     3061 tgacgtaatt attagaaagt gttctcaacg tttcttgcat agaatcataa tctctttttg
     3121 ataatacaac aactgtatct tctacatctt tacttgttac aataagtgtt tcagcatcct
     3181 cattaacttg tttcatataa ctacgtaaat tttggcggaa atttgaataa gctactgctt
     3241 ccattccttt caccccttat ttatttctat aaaacaattg tacattatat tgtacaatta
     3301 agcaaatgat taatattcta acttcttata attaaaagtg ctatcacttt tagtttttag
     3361 gataaaaagt gacgtcactt tttgagtgct ggtacctcaa atcctgcttt ttcttgggca
     3421 atcaataaat ctaacgcaat acacataatc ctggattcac ttttgccgta cttctgcttt
     3481 agttgctgca attctttttg tctcttttca tccaacgtaa cgggtaaccg tttatttcct
     3541 attgcaattg gcattagtac tctctccttt catcgaaata accctagtct atggacagtt
     3601 ttccctttga tatgtaacgg tgaacagttg ttctactttt gtttgttagt cttgatgctt
     3661 cactgataga tacaagagcc ataagaacct cagatccttc cgtatttagc cagtatgttc
     3721 tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg aaccattgag atcatactta
     3781 ctttgcatgt cactcaaaaa ttttgcctca aaactggtga gctgaatttt tgcagttaaa
     3841 gcatcgtgta gtgtttttct tagtccgtta tgtaggtagg aatctgatgt aatggttgtt
     3901 ggtattttgt caccattcat ttttatctgg ttgttctcaa gttcggttac gagatccatt
     3961 tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg ggcggcctcg cttatcaacc
     4021 accaatttca tattgctgta agtgtttaaa tctttactta ttggtttcaa aacccattgg
     4081 ttaagccttt taaactcatg gtagttattt tcaagcatta acatgaactt aaattcatca
     4141 aggctaatct ctatatttgc cttgtgagtt ttcttttgtg ttagttcttt taataaccac
     4201 tcataaatcc tcatagagta tttgttttca aaagacttaa catgttccag attatatttt
     4261 atgaattttt ttaactggaa aagataaggc aatatctctt cactaaaaac taattctaat
     4321 ttttcgcttg agaacttggc atagtttgtc cactggaaaa tctcaaagcc tttaaccaaa
     4381 ggattcctga tttccacagt tctcgtcatc agctctctgg ttgctttagc taatacacca
     4441 taagcatttt ccctactgat gttcatcatc tgagcgtatt ggttataagt gaacgatacc
     4501 gtccgttctt tccttgtagg gttttcaatc gtggggttga gtagtgccac acagcataaa
     4561 attagcttgg tttcatgctc cgttaagtca tagcgactaa tcgctagttc atttgctttg
     4621 aaaacaacta attcagacat acatctcaat tggtctaggt gattttaatc actataccaa
     4681 ttgagatggg ctagtcaatg ataattacta gtccttttcc tttgagttgt gggtatctgt
     4741 aaattctgct agacctttgc tggaaaactt gtaaattctg ctagaccctc tgtaaattcc
     4801 gctagacctt tgtgtgtttt ttttgtttat attcaagtgg ttataattta tagaataaag
     4861 aaagaataaa aaaagataaa aagaatagat cccagccctg tgtataactc actactttag
     4921 tcagttccgc agtattacaa aaggatgtcg
//
