LOCUS       sequence-418186-        3908 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU2-b backbone with kanamycin resistance and pSC101-var2 ORI. 
            amilCP dropout vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1670 to 1704)
  AUTHORS   Shapiro M
  TITLE     EcoFlex MoClo Extension, Volume I
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 3908)
  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            "unpublished"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_Sv2K_amilCP
FEATURES             Location/Qualifiers
     source          1..3908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(556..1350)
                     /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(667..686)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1277..1296
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1582..1600
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1604..1647)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1670..1704
                     /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             1713..1724
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2397..2468
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2491..2585)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2686..3636)
                     /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      3684..3906
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 tactttagtc agttccgcag tattacaaaa ggatgtcgca aacgctgttt gctcctctac
       61 aaaacagacc ttaaaaccct aaaggcttaa gtagcaccct cgcaagctcg gttgcggccg
      121 caatcgggca aatcgctgaa tattcctttt gtctccgacc atcaggcacc tgagtcgctg
      181 tctttttcgt gacattcagt tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg
      241 gcactacagg cgccttttat ggattcatgc aaggaaacta cccataatac aagaaaagcc
      301 cgtcacgggc ttccaggcgt ttattggcgg gtctgctatg tggtgctatc tgactttttg
      361 ctgttcagca gttcctgccc tctgattttc cagtctgacc acttcggatt atcccgtgac
      421 aggtcattca gactggctaa tgcacccagt aaggcagcgg tatcatcaac aagcaaagaa
      481 aaaaggcgac agattactct gtcgcctttt ttcctgactc ataaccaagc caattctcga
      541 accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga
      601 atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc
      661 ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg
      721 gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc
      781 atcgccgtgg gtcacgacga gatcctcgcc gtcgggcata cgcgccttga gcctggcgaa
      841 cagttcggct ggcgcgagcc cctgatgatc ttcgtccaga tcatcctgat cgacaagacc
      901 ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca
      961 ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc
     1021 ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca
     1081 gtcccttccc gcttcagtga caacgtcgag cacagccgcg caaggaacgc ccgtcgtggc
     1141 cagccacgat agccgcgctg cctcgtcttg cagttcattc agggcaccgg acaggtcggt
     1201 cttgacaaaa agaaccgggc gaccctgcgc tgacagccgg aacacggcgg catcagagca
     1261 gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag ccgccggaga
     1321 acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc
     1381 agatcttgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc agtttacttt
     1441 gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccggca ccccttgtat
     1501 tactgtttat gtaagcagac agttttattg ttgccacctg acgtctaaga aaccattatt
     1561 atcatgacat taacctataa aaataggcgt atcacgaggc agaatttcag ataaaaaaaa
     1621 tccttagctt tcgctaagga tgatttcggt ctcactatat ctagagacgt tgacggctag
     1681 ctcagtccta ggtacagtgc tagctactag agaaagagga gaaatactag atgagtgtga
     1741 tcgctaaaca aatgacctac aaggtttata tgtcaggcac ggtcaatgga cactactttg
     1801 aggtcgaagg cgatggaaaa ggtaagccct acgaggggga gcagacggta aagctcactg
     1861 tcaccaaggg cggacctctg ccatttgctt gggatatttt atcaccacag tgtcagtacg
     1921 gaagcatacc attcaccaag taccctgaag atatccctga ctatgtaaag cagtcattcc
     1981 cggagggcta tacatgggag aggatcatga actttgaaga tggtgcagtg tgtactgtca
     2041 gcaatgattc cagcatccaa ggcaactgtt tcatctacca tgtcaagttc tctggtttga
     2101 actttcctcc caatggacct gtcatgcaga agaaaacaca gggctgggaa cccaacactg
     2161 agcgtctgtt tgcacgagat ggaatgctgc taggaaacaa ctttatggct ctgaagttag
     2221 aaggaggcgg tcactatttg tgtgaattta aaactactta caaggcaaag aagcctgtga
     2281 agatgccagg gtatcactat gttgaccgca aactggatgt aaccaatcac aacaaggatt
     2341 acacttcggt tgagcagtgt gaaatttcca ttgcacgcaa acctgtggtc gcctaacaaa
     2401 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     2461 acgctctccg tctcagaagg tacagagacc attctcacca ataaaaaacg cccggcggca
     2521 accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag
     2581 gagtctatgg acagttttcc ctttgatatg taacggtgaa cagttgttct acttttgttt
     2641 gttagtcttg atgcttcact gatagataca agagccataa gaacctcaga tccttccgta
     2701 tttagccagt atgttctcta gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc
     2761 attgagatca tacttacttt gcatgtcact caaaaatttt gcctcaaaac tggtgagctg
     2821 aatttttgca gttaaagcat cgtgtagtgt ttttcttagt ccgttatgta ggtaggaatc
     2881 tgatgtaatg gttgttggta ttttgtcacc attcattttt atctggttgt tctcaagttc
     2941 ggttacgaga tccatttgtc tatctagttc aacttggaaa atcaacgtat cagtcgggcg
     3001 gcctcgctta tcaaccacca atttcatatt gctgtaagtg tttaaatctt tacttattgg
     3061 tttcaaaacc cattggttaa gccttttaaa ctcatggtag ttattttcaa gcattaacat
     3121 gaacttaaat tcatcaaggc taatctctat atttgccttg tgagttttct tttgtgttag
     3181 ttcttttaat aaccactcat aaatcctcat agagtatttg ttttcaaaag acttaacatg
     3241 ttccagatta tattttatga atttttttaa ctggaaaaga taaggcaata tctcttcact
     3301 aaaaactaat tctaattttt cgcttgagaa cttggcatag tttgtccact ggaaaatcca
     3361 aaagccttta accaaaggat tcctgatttc cacagttctc gtcatcagct ctctggttgc
     3421 tttagctaat acaccataag cattttccct actgatgttc atcatctgag cgtattggtt
     3481 ataagtgaac gataccgtcc gttctttcct tgtagggttt tcaatcgtgg ggttgagtag
     3541 tgccacacag cataaaatta gcttggtttc atgctccgtt aagtcatagc gactaatcgc
     3601 tagttcattt gctttgaaaa caactaattc agacatacat ctcaattggt ctaggtgatt
     3661 ttaatcacta taccaattga gatgggctag tcaatgataa ttactagtcc ttttcctttg
     3721 agttgtgggt atctgtaaat tctgctagac ctttgctgga aaacttgtaa attctgctag
     3781 accctctgta aattccgcta gacctttgtg tgtttttttt gtttatattc aagtggttat
     3841 aatttataga ataaagaaag aataaaaaaa gataaaaaga atagatccca gccctgtgta
     3901 taactcac
//
