LOCUS       sequence-413884-        3910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU2-b backbone with kanamycin resistance and pSC101 (WT) ORI. 
            amilCP dropout vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1674 to 1708)
  AUTHORS   Shapiro M
  TITLE     EcoFlex MoClo Extension, Volume I
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 3910)
  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            "unpublished"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_SK_amilCP
FEATURES             Location/Qualifiers
     source          1..3910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(560..1354)
                     /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(671..690)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1281..1300
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1586..1604
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1608..1651)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1674..1708
                     /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             1717..1728
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2401..2472
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2495..2589)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2690..3640)
                     /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      3688..3910
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 actactttag tcagttccgc agtattacaa aaggatgtcg caaacgctgt ttgctcctct
       61 acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc ctcgcaagct cggttgcggc
      121 cgcaatcggg caaatcgctg aatattcctt ttgtctccga ccatcaggca cctgagtcgc
      181 tgtctttttc gtgacattca gttcgctgcg ctcacggctc tggcagtgaa tgggggtaaa
      241 tggcactaca ggcgcctttt atggattcat gcaaggaaac tacccataat acaagaaaag
      301 cccgtcacgg gcttctcagg gcgttttatg gcgggtctgc tatgtggtgc tatctgactt
      361 tttgctgttc agcagttcct gccctctgat tttccagtct gaccacttcg gattatcccg
      421 tgacaggtca ttcagactgg ctaatgcacc cagtaaggca gcggtatcat caacaagcaa
      481 agaaaaaagg cgacagatta ctctgtcgcc ttttttcctg actcataacc aagccaattc
      541 tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct
      601 gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa
      661 gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca
      721 gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc
      781 aggcatcgcc gtgggtcacg acgagatcct cgccgtcggg catacgcgcc ttgagcctgg
      841 cgaacagttc ggctggcgcg agcccctgat gatcttcgtc cagatcatcc tgatcgacaa
      901 gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg
      961 ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt
     1021 tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca
     1081 gccagtccct tcccgcttca gtgacaacgt cgagcacagc cgcgcaagga acgcccgtcg
     1141 tggccagcca cgatagccgc gctgcctcgt cttgcagttc attcagggca ccggacaggt
     1201 cggtcttgac aaaaagaacc gggcgaccct gcgctgacag ccggaacacg gcggcatcag
     1261 agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc caagccgccg
     1321 gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt
     1381 gatcagatct tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta
     1441 ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc ggcacccctt
     1501 gtattactgt ttatgtaagc agacagtttt attgttgcca cctgacgtct aagaaaccat
     1561 tattatcatg acattaacct ataaaaatag gcgtatcacg aggcagaatt tcagataaaa
     1621 aaaatcctta gctttcgcta aggatgattt cggtctcact atatctagag acgttgacgg
     1681 ctagctcagt cctaggtaca gtgctagcta ctagagaaag aggagaaata ctagatgagt
     1741 gtgatcgcta aacaaatgac ctacaaggtt tatatgtcag gcacggtcaa tggacactac
     1801 tttgaggtcg aaggcgatgg aaaaggtaag ccctacgagg gggagcagac ggtaaagctc
     1861 actgtcacca agggcggacc tctgccattt gcttgggata ttttatcacc acagtgtcag
     1921 tacggaagca taccattcac caagtaccct gaagatatcc ctgactatgt aaagcagtca
     1981 ttcccggagg gctatacatg ggagaggatc atgaactttg aagatggtgc agtgtgtact
     2041 gtcagcaatg attccagcat ccaaggcaac tgtttcatct accatgtcaa gttctctggt
     2101 ttgaactttc ctcccaatgg acctgtcatg cagaagaaaa cacagggctg ggaacccaac
     2161 actgagcgtc tgtttgcacg agatggaatg ctgctaggaa acaactttat ggctctgaag
     2221 ttagaaggag gcggtcacta tttgtgtgaa tttaaaacta cttacaaggc aaagaagcct
     2281 gtgaagatgc cagggtatca ctatgttgac cgcaaactgg atgtaaccaa tcacaacaag
     2341 gattacactt cggttgagca gtgtgaaatt tccattgcac gcaaacctgt ggtcgcctaa
     2401 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     2461 gtgaacgctc tccgtctcag aaggtacaga gaccattctc accaataaaa aacgcccggc
     2521 ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca
     2581 acaggagtct atggacagtt ttccctttga tatgtaacgg tgaacagttg ttctactttt
     2641 gtttgttagt cttgatgctt cactgataga tacaagagcc ataagaacct cagatccttc
     2701 cgtatttagc cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg
     2761 aaccattgag atcatactta ctttgcatgt cactcaaaaa ttttgcctca aaactggtga
     2821 gctgaatttt tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta tgtaggtagg
     2881 aatctgatgt aatggttgtt ggtattttgt caccattcat ttttatctgg ttgttctcaa
     2941 gttcggttac gagatccatt tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg
     3001 ggcggcctcg cttatcaacc accaatttca tattgctgta agtgtttaaa tctttactta
     3061 ttggtttcaa aacccattgg ttaagccttt taaactcatg gtagttattt tcaagcatta
     3121 acatgaactt aaattcatca aggctaatct ctatatttgc cttgtgagtt ttcttttgtg
     3181 ttagttcttt taataaccac tcataaatcc tcatagagta tttgttttca aaagacttaa
     3241 catgttccag attatatttt atgaattttt ttaactggaa aagataaggc aatatctctt
     3301 cactaaaaac taattctaat ttttcgcttg agaacttggc atagtttgtc cactggaaaa
     3361 tctcaaagcc tttaaccaaa ggattcctga tttccacagt tctcgtcatc agctctctgg
     3421 ttgctttagc taatacacca taagcatttt ccctactgat gttcatcatc tgagcgtatt
     3481 ggttataagt gaacgatacc gtccgttctt tccttgtagg gttttcaatc gtggggttga
     3541 gtagtgccac acagcataaa attagcttgg tttcatgctc cgttaagtca tagcgactaa
     3601 tcgctagttc atttgctttg aaaacaacta attcagacat acatctcaat tggtctaggt
     3661 gattttaatc actataccaa ttgagatggg ctagtcaatg ataattacta gtccttttcc
     3721 tttgagttgt gggtatctgt aaattctgct agacctttgc tggaaaactt gtaaattctg
     3781 ctagaccctc tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat attcaagtgg
     3841 ttataattta tagaataaag aaagaataaa aaaagataaa aagaatagat cccagccctg
     3901 tgtataactc
//
