LOCUS       sequence-435736-        5145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 pAHP6 promoter, GG0 and GG1 BsaI sites for golden gate 
            assembly..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5145)
  TITLE     Nemhauser Lab Plasmids 2024
REFERENCE   2  (bases 1 to 5145)
  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"
COMMENT     Sequence Label: L0I84 - GG0-pAHP6-GG1
FEATURES             Location/Qualifiers
     source          1..5145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      110..119
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(319..1113)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(319..1113)
                     /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
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(430..449)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1040..1059
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(1462..1764)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(1564..1583)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     1887..1909
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1901..1918
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1902..1918
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1925..1944
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1925..1943
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3756..3772)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3756..3772)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3769..3791)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3780..3796
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3804..3810,3811..3828,3829..3834))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3849..3870
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3987..4004)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4158..4746)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4238..4257)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4884..5145,1..113))
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
ORIGIN
        1 aagtcgtcct ccacgaagtc ccgggagaac ccgagccggt cggtccagaa ctcgaccgct
       61 ccggcgacgt cgcgcgcggt gagcaccgga acggcactgg tcaacttggc catggtggcc
      121 ctcctcacgt gctattattg aagcatttat cagggttatt gtctcatgag cggatacata
      181 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
      241 ccacctgatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa
      301 attgtaagcg ttaataattc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
      361 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
      421 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag
      481 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
      541 ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct tgagcctggc
      601 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
      661 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
      721 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
      781 ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag
      841 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
      901 ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc
      961 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     1021 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     1081 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg
     1141 atcagagctt gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac
     1201 tttgcagggc ttcccaacct taccagaggg cgccccagct ggcaattccg gttcgcttgc
     1261 tgtccataaa accgcccagt ctagctatcg ccatgtaagc ccactgcaag ctacctgctt
     1321 tctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatccgggg
     1381 tcagcaccgt ttctgcggac tggctttcta cgtgaaaagg atctaggtga agatcctttt
     1441 tgataatctc atgcctgaca tttatattcc ccagaacatc aggttaatgg cgtttttgat
     1501 gtcattttcg cggtggctga gatcagccac ttcttccccg ataacggaga ccggcacact
     1561 ggccatatcg gtggtcatca tgcgccagct ttcatccccg atatgcacca ccgggtaaag
     1621 ttcacgggag actttatctg acagcagacg tgcactggcc agggggatca ccatccgtcg
     1681 ccccggcgtg tcaataatat cactctgtac atccacaaac agacgataac ggctctctct
     1741 tttataggtg taaaccttaa actgccgtac gtataggctg cgcaactgtt gggaagggcg
     1801 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     1861 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     1921 attgtaatac gactcactat agggcgaatt gggccctcta gatgcatgct cgagcggccg
     1981 ccagtgtgat ggatatctgc agaattcagg acgttgatgg tctcacattc aacggcacac
     2041 ccgtcttgtc ccgttttccg cttcaatctt tcctctttct atatatttgt ttttactaat
     2101 gtgaacaggt cttgcttcaa attctagcta gagaaggctt ataattcgat tttgggtttg
     2161 ttgagaggta tttataattg tatagacatg actacatgag atgaggtgac ccacaattgt
     2221 attcaaattt tctaagagtc aggttacctt tttcccccac ttttaagtga aagaatcgag
     2281 gagggtgtgg ggaataaacg ggaccgtcac aggaaaaagc actctacaag tattgtacga
     2341 ccatattaat acactctgtg accggctaca gccgccagcc ggttaccccg cgtgactctg
     2401 ccaatagcgg cgttcttcct cccgacaacc ggcgtaccct gtcagaccag ctggtcaccc
     2461 ttcccgcagc ctctacattt acatatttaa cttaatacat ctgccttaaa ttaaatcaac
     2521 aacttttaca ccaataatat gtactttcta tctttaattt ctcactgttg aaattaatct
     2581 cttttctttt atcaggtctg attaagatga ttatttggtc tgtcaagtta taatcatata
     2641 acagtaatgt acattagatt atctcataat taatccactc ttgatttaac acaagaattg
     2701 tttttttttt tttttttttt taaatgtgag catgcattag tttctgagac tgtaaaagaa
     2761 aagcccacac tttgttgctc cctcgtatga cgcgcgactt tcccactaac tttgcagtga
     2821 gtgttcgcta ttctctatgc ttcaaattca atccatcctt tctccatttc tatctcaaaa
     2881 ggcctttctc tttctcgatc taatcaatta atcttcttct ttttctttct tataaggaaa
     2941 gctatacaca agaagaatac aagaaacaca agcacaacac agaaacatgc gtaaattgtt
     3001 tcaccgaata aagtaaaaaa ttatcaatag ttaaacacaa aagattacaa gtcctcttaa
     3061 ctatttttaa cagaaaatgt atgtaccaac acctctggtt acgtgatttt ctcataactt
     3121 tggtgattgg tatattggta tcacatcgct atatatgaac catgacagta cataaataaa
     3181 taatcatatg gcacgtataa gaagcctcta ataatcttcc ttaattagtc tcctggaacc
     3241 agaaagtata cataattata aaaagcagcg gtaactaacg aagatttata atatttgaac
     3301 acaaaaaagc gtgctcatat gtaattattg tgagctagta tagaatgata tatgaaaata
     3361 gctaaatatt attgtaaatg cggattaaaa atgactgaaa agaagaaaga gcgtgcttgg
     3421 aaacggaaca aacaactagc tagtattaaa catatacttt ttcatgttat cagagtaagt
     3481 agtaaagtgg agcgcataat gagtggatgc ttttttaagg ttatgttacc aattaaaact
     3541 agtgattaaa ttatgattat ggaattatta tatacgaaaa ctgaaattgt ctcattcgta
     3601 tgtcatgctt ctttgcgccc cgtgctcctg agaccacaga gtgcctgaat tccagcacac
     3661 tggcggccgt tactagtgga tccgagctcg gtaccaagct tgatgcatag cttgagtatt
     3721 ctaacgcgtc acctaaatag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3781 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     3841 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3901 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3961 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4021 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4081 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4141 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4201 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4261 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4321 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4381 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4441 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4501 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4561 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4621 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4681 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4741 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4801 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4861 taaaaatgaa gttttagcac gtgtcagtcc tgctcctcgg ccacgaagtg cacgcagttg
     4921 ccggccgggt cgcgcagggc gaactcccgc ccccacggct gctcgccgat ctcggtcatg
     4981 gccggcccgg aggcgtcccg gaagttcgtg gacacgacct ccgaccactc ggcgtacagc
     5041 tcgtccaggc cgcgcaccca cacccaggcc agggtgttgt ccggcaccac ctggtcctgg
     5101 accgcgctga tgaacagggt cacgtcgtcc cggaccacac cggcg
//
