LOCUS       sequence-418156-        3910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU1-A backbone with kanamycin resistance and pSC101-var6 ORI. 
            amilCP dropout vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3910)
  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  JOURNAL   unpublished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            "unpublished"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: pTU1-A_Sv6K_amilCP
FEATURES             Location/Qualifiers
     source          1..3910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(576..1370)
                     /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(687..706)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1297..1316
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1602..1620
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1624..1667)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1690..1724
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             1733..1744
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2417..2488
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2511..2605)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2706..3656)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTDYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      join(3704..3910,1..16)
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 gccctgtgta taactcacta ctttagtcag ttccgcagta ttacaaaagg atgtcgcaaa
       61 cgctgtttgc tcctctacaa aacagacctt aaaaccctaa aggcttaagt agcaccctcg
      121 caagctcggt tgcggccgca atcgggcaaa tcgctgaata ttccttttgt ctccgaccat
      181 caggcacctg agtcgctgtc tttttcgtga cattcagttc gctgcgctca cggctctggc
      241 agtgaatggg ggtaaatggc actacaggcg ccttttatgg attcatgcaa ggaaactacc
      301 cataatacaa gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg gtctgctatg
      361 tggtgctatc tgactttttg ctgttcagca gttcctgccc tctgattttc cagtctgacc
      421 acttcggatt atcccgtgac aggtcattca gactggctaa tgcacccagt aaggcagcgg
      481 tatcatcaac aagcaaagaa aaaaggcgac agattactct gtcgcctttt ttcctgactc
      541 ataaccaagc caattctcga accccagagt cccgctcaga agaactcgtc aagaaggcga
      601 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
      661 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
      721 cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
      781 atgatattcg gcaagcaggc atcgccgtgg gtcacgacga gatcctcgcc gtcgggcata
      841 cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgatc ttcgtccaga
      901 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
      961 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     1021 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
     1081 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagccgcg
     1141 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc
     1201 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gaccctgcgc tgacagccgg
     1261 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     1321 tccacccaag ccgccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
     1381 cctcatcctg tctcttgatc agatcttgat cccctgcgcc atcagatcct tggcggcaag
     1441 aaagccatcc agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc
     1501 aattccggca ccccttgtat tactgtttat gtaagcagac agttttattg ttgccacctg
     1561 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     1621 agaatttcag ataaaaaaaa tccttagctt tcgctaagga tgatttccgt ctcaatctct
     1681 atagagacct ttacagctag ctcagtccta ggtattatgc tagctactag agaaagagga
     1741 gaaatactag ataagtgtga tcgctaaaca aatgacctac aaggtttata tgtcaggcac
     1801 ggtcaatgga cactactttg aggtcgaagg cgatggaaaa ggtaagccct acgaggggga
     1861 gcagacggta aagctcactg tcaccaaggg cggacctctg ccatttgctt gggatatttt
     1921 atcaccacag tgtcagtacg gaagcatacc attcaccaag taccctgaag atatccctga
     1981 ctatgtaaag cagtcattcc cggagggcta tacatgggag aggatcatga actttgaaga
     2041 tggtgcagtg tgtactgtca gcaatgattc cagcatccaa ggcaactgtt tcatctacca
     2101 tgtcaagttc tctggtttga actttcctcc caatggacct gtcatgcaga agaaaacaca
     2161 gggctgggaa cccaacactg agcgtctgtt tgcacgagat ggaatgctgc taggaaacaa
     2221 ctttatggct ctgaagttag aaggaggcgg tcactatttg tgtgaattta aaactactta
     2281 caaggcaaag aagcctgtga agatgccagg gtatcactat gttgaccgca aactggatgt
     2341 aaccaatcac aacaaggatt acacttcggt tgagcagtgt gaaatttcca ttgcacgcaa
     2401 acctgtggtc gcctaacaaa taaaacgaaa ggatcagtcg aaagactggg cctttcgttt
     2461 tatctgttgt ttgtcggtga acgctctcgg tctcatgttt gccagagacg attctcacca
     2521 ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
     2581 attactggat ctatcaacag gagtctatgg acagttttcc ctttgatatg taacggtgaa
     2641 cagttgttct acttttgttt gttagtcttg atgcttcact gatagataca agagccataa
     2701 gaacctcaga tccttccgta tttagccagt atgttctcta gtgtggttcg ttgtttttgc
     2761 gtgagccatg agaacgaacc attgagatca tacttacttt gcatgtcact caaaaatttt
     2821 gcctcaaaac tggtgagctg aatttttgca gttaaagcat cgtgtagtgt ttttcttagt
     2881 ccgttatgta ggtaggaatc tgatgtaatg gttgttggta ttttgtcacc attcattttt
     2941 atctggttgt tctcaagttc ggttacgaga tccatttgtc tatctagttc aacttggaaa
     3001 atcaacgtat cagtcgggcg gcctcgctta tcaaccacca atttcatatt gctgtaagtg
     3061 tttaaatctt tacttattgg tttcaaaacc cattggttaa gccttttaaa ctcatggtag
     3121 ttattttcaa gcattaacat gaacttaaat tcatcaaggc taatctctat atttgccttg
     3181 tgagttttct tttgtgttag ttcttttaat aaccactcat aaatcctcat agagtatttg
     3241 ttttcaaaag acttaacatg ttccagatta tattttatga atttttttaa ctggaaaaga
     3301 taaggcaata tctcttcact aaaaactaat tctaattttt cgcttgagaa cttggcatag
     3361 tctgtccact ggaaaatctc aaagccttta accaaaggat tcctgatttc cacagttctc
     3421 gtcatcagct ctctggttgc tttagctaat acaccataag cattttccct actgatgttc
     3481 atcatctgag cgtattggtt ataagtgaac gataccgtcc gttctttcct tgtagggttt
     3541 tcaatcgtgg ggttgagtag tgccacacag cataaaatta gcttggtttc atgctccgtt
     3601 aagtcatagc gactaatcgc tagttcattt gctttgaaaa caactaattc agacatacat
     3661 ctcaattggt ctaggtgatt ttaatcacta taccaattga gatgggctag tcaatgataa
     3721 ttactagtcc ttttcctttg agttgtgggt atctgtaaat tctgctagac ctttgctgga
     3781 aaacttgtaa attctgctag accctctgta aattccgcta gacctttgtg tgtttttttt
     3841 gtttatattc aagtggttat aatttataga ataaagaaag aataaaaaaa gataaaaaga
     3901 atagatccca
//
