LOCUS       sequence-413880-        4946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU2-a backbone with kanamycin resistance, pSC101-var4 ORI, and 
            Axe-Txe plasmid stability cassette. amilCP dropout vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4946)
  AUTHORS   Shapiro M
  TITLE     EcoFlex MoClo Extension, Volume I
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 4946)
  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            "unpublished"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_Sv4KA_amilCP
FEATURES             Location/Qualifiers
     source          1..4946
                     /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(1488..1531)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1554..1588
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             1597..1608
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2281..2352
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2375..2469)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3686..4636)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFGIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      4684..4906
                     /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 agacagtatt tcagataaaa
     1501 aaaatcctta gctttcgcta aggatgattt cggtctcact atatctagag acgtttacag
     1561 ctagctcagt cctaggtatt atgctagcta ctagagaaag aggagaaata ctagatgagt
     1621 gtgatcgcta aacaaatgac ctacaaggtt tatatgtcag gcacggtcaa tggacactac
     1681 tttgaggtcg aaggcgatgg aaaaggtaag ccctacgagg gggagcagac ggtaaagctc
     1741 actgtcacca agggcggacc tctgccattt gcttgggata ttttatcacc acagtgtcag
     1801 tacggaagca taccattcac caagtaccct gaagatatcc ctgactatgt aaagcagtca
     1861 ttcccggagg gctatacatg ggagaggatc atgaactttg aagatggtgc agtgtgtact
     1921 gtcagcaatg attccagcat ccaaggcaac tgtttcatct accatgtcaa gttctctggt
     1981 ttgaactttc ctcccaatgg acctgtcatg cagaagaaaa cacagggctg ggaacccaac
     2041 actgagcgtc tgtttgcacg agatggaatg ctgctaggaa acaactttat ggctctgaag
     2101 ttagaaggag gcggtcacta tttgtgtgaa tttaaaacta cttacaaggc aaagaagcct
     2161 gtgaagatgc cagggtatca ctatgttgac cgcaaactgg atgtaaccaa tcacaacaag
     2221 gattacactt cggttgagca gtgtgaaatt tccattgcac gcaaacctgt ggtcgcctaa
     2281 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     2341 gtgaacgctc tccgtctcac cgggtacaga gaccattctc accaataaaa aacgcccggc
     2401 ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca
     2461 acaggagtca agcgtcatcc ttagtaacta acatcccaca ctacaatagt attataacat
     2521 atctattcaa cttcaaattg ttgaaattta agccatttat atttactttt cctcaacttt
     2581 agaaatgatt ttactaacct tatatccgta atacgcgtaa caaacacatt cattaataac
     2641 aatcaacact aagctatatt agctttttaa atgactaggg taaataaaat aaggatcaaa
     2701 aacttttaag tttctgaccc tttccttact tccgattggt taatagtgat cttttgcaga
     2761 ataaataaat atcgtttcat tttcaactct atatatcagt ctatgttcat ctgtaattct
     2821 tctggaccat tttccagata aatcatgctt taatggctca ggttttccta atccagcaaa
     2881 gggggaacga tcgatatctt ttattaactt gttaatcttt tttatattgc ttttgtttcc
     2941 ttgctcatgc caataaagat aatcatccca agcatcatca gaccaagcct taatcatcag
     3001 attcaacctc gattaagtca tgtgttttaa atgcaccttt ggagaattgt tcatctcctc
     3061 gacgaatttt ttccatgacg taattattag aaagtgttct caacgtttct tgcatagaat
     3121 cataatctct ttttgataat acaacaactg tatcttctac atctttactt gttacaataa
     3181 gtgtttcagc atcctcatta acttgtttca tataactacg taaattttgg cggaaatttg
     3241 aataagctac tgcttccatt cctttcaccc cttatttatt tctataaaac aattgtacat
     3301 tatattgtac aattaagcaa atgattaata ttctaacttc ttataattaa aagtgctatc
     3361 acttttagtt tttaggataa aaagtgacgt cactttttga gtgctggtac ctcaaatcct
     3421 gctttttctt gggcaatcaa taaatctaac gcaatacaca taatcctgga ttcacttttg
     3481 ccgtacttct gctttagttg ctgcaattct ttttgtctct tttcatccaa cgtaacgggt
     3541 aaccgtttat ttcctattgc aattggcatt agtactctcc tagtctatgg acagttttcc
     3601 ctttgatatg taacggtgaa cagttgttct acttttgttt gttagtcttg atgcttcact
     3661 gatagataca agagccataa gaacctcaga tccttccgta tttagccagt atgttctcta
     3721 gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc attgagatca tacttacttt
     3781 gcatgtcact caaaaatttt gcctcaaaac tggtgagctg aatttttgca gttaaagcat
     3841 cgtgtagtgt ttttcttagt ccgttatgta ggtaggaatc tgatgtaatg gttgttggta
     3901 ttttgtcacc attcattttt atctggttgt tctcaagttc ggttacgaga tccatttgtc
     3961 tatctagttc aacttggaaa atcaacgtat cagtcgggcg gcctcgctta tcaaccacca
     4021 atttcatatt gctgtaagtg tttaaatctt tacttattgg tttcaaaacc cattggttaa
     4081 gccttttaaa ctcatggtag ttattttcaa gcattaacat gaacttaaat tcatcaaggc
     4141 taatctctat atttgccttg tgagttttct tttgtgttag ttcttttaat aaccactcat
     4201 aaatcctcat agagtatttg ttttcaaaag acttaacatg ttccagatta tattttatga
     4261 atttttttaa ctggaaaaga taaggcaata tctcttcact aaaaactaat tctaattttt
     4321 cgcttgagaa cttggcatag tttgtccact ggaaaatccc aaagccttta accaaaggat
     4381 tcctgatttc cacagttctc gtcatcagct ctctggttgc tttagctaat acaccataag
     4441 cattttccct actgatgttc atcatctgag cgtattggtt ataagtgaac gataccgtcc
     4501 gttctttcct tgtagggttt tcaatcgtgg ggttgagtag tgccacacag cataaaatta
     4561 gcttggtttc atgctccgtt aagtcatagc gactaatcgc tagttcattt gctttgaaaa
     4621 caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga
     4681 gatgggctag tcaatgataa ttactagtcc ttttcctttg agttgtgggt atctgtaaat
     4741 tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta
     4801 gacctttgtg tgtttttttt gtttatattc aagtggttat aatttataga ataaagaaag
     4861 aataaaaaaa gataaaaaga atagatccca gccctgtgta taactcacta ctttagtcag
     4921 ttccgcagta ttacaaaagg atgtcg
//
