LOCUS       sequence-418160-        3908 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU1-A 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 1664 to 1698)
  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  JOURNAL   unpublished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            "unpublished"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: pTU1-A_Sv2K_amilCP
FEATURES             Location/Qualifiers
     source          1..3908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(550..1344)
                     /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(661..680)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1271..1290
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1576..1594
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1598..1641)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1664..1698
                     /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             1707..1718
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2391..2462
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2485..2579)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2680..3630)
                     /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      3678..3900
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 agtcagttcc gcagtattac aaaaggatgt cgcaaacgct gtttgctcct ctacaaaaca
       61 gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag ctcggttgcg gccgcaatcg
      121 ggcaaatcgc tgaatattcc ttttgtctcc gaccatcagg cacctgagtc gctgtctttt
      181 tcgtgacatt cagttcgctg cgctcacggc tctggcagtg aatgggggta aatggcacta
      241 caggcgcctt ttatggattc atgcaaggaa actacccata atacaagaaa agcccgtcac
      301 gggcttccag gcgtttattg gcgggtctgc tatgtggtgc tatctgactt tttgctgttc
      361 agcagttcct gccctctgat tttccagtct gaccacttcg gattatcccg tgacaggtca
      421 ttcagactgg ctaatgcacc cagtaaggca gcggtatcat caacaagcaa agaaaaaagg
      481 cgacagatta ctctgtcgcc ttttttcctg actcataacc aagccaattc tcgaacccca
      541 gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg
      601 agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc
      661 aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca
      721 gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc
      781 gtgggtcacg acgagatcct cgccgtcggg catacgcgcc ttgagcctgg cgaacagttc
      841 ggctggcgcg agcccctgat gatcttcgtc cagatcatcc tgatcgacaa gaccggcttc
      901 catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc
      961 cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg
     1021 agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct
     1081 tcccgcttca gtgacaacgt cgagcacagc cgcgcaagga acgcccgtcg tggccagcca
     1141 cgatagccgc gctgcctcgt cttgcagttc attcagggca ccggacaggt cggtcttgac
     1201 aaaaagaacc gggcgaccct gcgctgacag ccggaacacg gcggcatcag agcagccgat
     1261 tgtctgttgt gcccagtcat agccgaatag cctctccacc caagccgccg gagaacctgc
     1321 gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcagatct
     1381 tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta ctttgcaggg
     1441 cttcccaacc ttaccagagg gcgccccagc tggcaattcc ggcacccctt gtattactgt
     1501 ttatgtaagc agacagtttt attgttgcca cctgacgtct aagaaaccat tattatcatg
     1561 acattaacct ataaaaatag gcgtatcacg aggcagaatt tcagataaaa aaaatcctta
     1621 gctttcgcta aggatgattt ccgtctcaat ctctatagag accttgacgg ctagctcagt
     1681 cctaggtaca gtgctagcta ctagagaaag aggagaaata ctagatgagt gtgatcgcta
     1741 aacaaatgac ctacaaggtt tatatgtcag gcacggtcaa tggacactac tttgaggtcg
     1801 aaggcgatgg aaaaggtaag ccctacgagg gggagcagac ggtaaagctc actgtcacca
     1861 agggcggacc tctgccattt gcttgggata ttttatcacc acagtgtcag tacggaagca
     1921 taccattcac caagtaccct gaagatatcc ctgactatgt aaagcagtca ttcccggagg
     1981 gctatacatg ggagaggatc atgaactttg aagatggtgc agtgtgtact gtcagcaatg
     2041 attccagcat ccaaggcaac tgtttcatct accatgtcaa gttctctggt ttgaactttc
     2101 ctcccaatgg acctgtcatg cagaagaaaa cacagggctg ggaacccaac actgagcgtc
     2161 tgtttgcacg agatggaatg ctgctaggaa acaactttat ggctctgaag ttagaaggag
     2221 gcggtcacta tttgtgtgaa tttaaaacta cttacaaggc aaagaagcct gtgaagatgc
     2281 cagggtatca ctatgttgac cgcaaactgg atgtaaccaa tcacaacaag gattacactt
     2341 cggttgagca gtgtgaaatt tccattgcac gcaaacctgt ggtcgcctaa caaataaaac
     2401 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     2461 tcggtctcat gtttgccaga gacgattctc accaataaaa aacgcccggc ggcaaccgag
     2521 cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca acaggagtct
     2581 atggacagtt ttccctttga tatgtaacgg tgaacagttg ttctactttt gtttgttagt
     2641 cttgatgctt cactgataga tacaagagcc ataagaacct cagatccttc cgtatttagc
     2701 cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg aaccattgag
     2761 atcatactta ctttgcatgt cactcaaaaa ttttgcctca aaactggtga gctgaatttt
     2821 tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta tgtaggtagg aatctgatgt
     2881 aatggttgtt ggtattttgt caccattcat ttttatctgg ttgttctcaa gttcggttac
     2941 gagatccatt tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg ggcggcctcg
     3001 cttatcaacc accaatttca tattgctgta agtgtttaaa tctttactta ttggtttcaa
     3061 aacccattgg ttaagccttt taaactcatg gtagttattt tcaagcatta acatgaactt
     3121 aaattcatca aggctaatct ctatatttgc cttgtgagtt ttcttttgtg ttagttcttt
     3181 taataaccac tcataaatcc tcatagagta tttgttttca aaagacttaa catgttccag
     3241 attatatttt atgaattttt ttaactggaa aagataaggc aatatctctt cactaaaaac
     3301 taattctaat ttttcgcttg agaacttggc atagtttgtc cactggaaaa tccaaaagcc
     3361 tttaaccaaa ggattcctga tttccacagt tctcgtcatc agctctctgg ttgctttagc
     3421 taatacacca taagcatttt ccctactgat gttcatcatc tgagcgtatt ggttataagt
     3481 gaacgatacc gtccgttctt tccttgtagg gttttcaatc gtggggttga gtagtgccac
     3541 acagcataaa attagcttgg tttcatgctc cgttaagtca tagcgactaa tcgctagttc
     3601 atttgctttg aaaacaacta attcagacat acatctcaat tggtctaggt gattttaatc
     3661 actataccaa ttgagatggg ctagtcaatg ataattacta gtccttttcc tttgagttgt
     3721 gggtatctgt aaattctgct agacctttgc tggaaaactt gtaaattctg ctagaccctc
     3781 tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat attcaagtgg ttataattta
     3841 tagaataaag aaagaataaa aaaagataaa aagaatagat cccagccctg tgtataactc
     3901 actacttt
//
