LOCUS       sequence-435863-        6494 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 pGATA23-Bxb1-tUBQ10 construct for assembly of two integrase 
            constructs, GG4 and GG6 BsaI sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6494)
  TITLE     Nemhauser Lab Plasmids 2024
REFERENCE   2  (bases 1 to 6494)
  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: L0I101- GG4-pGATA23-Bxb1-tUBQ10-GG6
FEATURES             Location/Qualifiers
     source          1..6494
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             826..2328
                     /codon_start=1
                     /transl_table=11
                     /gene="Bxb1"
                     /product="Bxb1 integrase"
                     /label=Bxb1 integrase
                     /note="Bxb1 large serine integrase from Mycobacterium 
                     bacteriophage Bxb1 (Ghosh et al. 2003)."
                     /protein_id="AFF61396.1"
                     /translation="MRALVVIRLSRVTDATTSPERQLESCQQLCAQRGWDVVGVAEDLD
                     VSGAVDPFDRKRRPNLARWLAFEEQPFDVIVAYRVDRLTRSIRHLQQLVHWAEDHKKLV
                     VSATEAHFDTTTPFAAVVIALMGTVAQMELEAIKERNRSAAHFNIRAGKYRGSLPPWGY
                     LPTRVDGEWRLVPDPVQRERILEVYHRVVDNHEPLHLVAHDLNRRGVLSPKDYFAQLQG
                     REPQGREWSATALKRSMISGAMLGYATLNGKTVRDDDGAPLVRAEPILTREQLEALRAE
                     LVKTSRAKPAVSTPSLLLRVLFCAVCGEPAYKFAGGGRKHPRYRCRSMGFPKHCGNGTV
                     AMAEWDAFCEEQVLDLLGDAERLEKVWVAGSDSAVELAEVNAELVDLTSLIGSPAYRAG
                     SPQREALDARIAALAARQEELEGLEARPSGWEWRETGQRFGDWWREQDTAAKNTWLRSM
                     NVRLTFDVRGGLTRTIDFGDLQEYEQHLRLGSVVERLHTGMS"
     primer_bind     complement(3049..3068)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3050..3068)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3075..3092)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3075..3091)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3084..3106)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             3229..3531
                     /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     3410..3429
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             3880..4674
                     /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             3880..4674
                     /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(3934..3953)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4544..4563
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      4874..4883
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4880..5254
                     /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"
     rep_origin      5392..5980
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5881..5900
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6134..6151
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6268..6289
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6304..6309,6310..6327,6328..6334)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6342..6358
                     /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)."
     primer_bind     6347..6369
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6366..6382
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6366..6382
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cactctgtgg tctcagcttc atgaataatc tttgcgtaca cctaaatttt atagtgaaaa
       61 atacataatt ttaaaacttt acttttgtag agatgcctaa aaaaaaatgg tttaccacat
      121 ggacattgtt atttgttcag atctccaacg tatagcaata tttgtgattt atagatatgt
      181 aatttggtgt tcaaagaatg tgtagaacaa ttattcttat gggtatgcca agaaaagtta
      241 tataacttta ttctatactg ctttcacgtt aaataatgta acattattaa tgttgtcctt
      301 ttacttttcc ttgaattgat ttgggaagaa gactattggc ttccttatca catcaaaaag
      361 tctatgagta agtgaaacat tggctggctt ttgcttcatt cctttgtctc tgtcttcatg
      421 tgaacaagtt tgcagaaatt tgtggtttct ttttttactt tcttttgtct aatacacaca
      481 ctctcaccac tattataaag aaaatccata gatcccggtc gtgcgtactg ccactttgct
      541 ttccacccga caatgatttg ggagatgttt tgttttgtgt catacatcta ccaaaaaagt
      601 cttaatcatt gttcatcaaa agagcgccca agggctcttt cgtcttctta ccacacgcac
      661 actagttcga gagggttttt aaggagctct taagcctttc ttgatgactc ataccacgaa
      721 caagaaccaa tcttgtcacc catataatct ccgtattcag agaattgaag agttcttgat
      781 cactactgtt gtatcaaaaa ggaactctaa gaactaagat tgaaaatgag agccctggta
      841 gtcatccgcc tgtcccgcgt caccgatgct acgacttcac cggagcgtca gctggagtct
      901 tgccagcagc tctgcgccca gcgcggctgg gacgtcgtcg gggtagcgga ggatctggac
      961 gtctccgggg cggtcgatcc gttcgaccgg aagcgcagac cgaacctggc ccggtggcta
     1021 gcgttcgagg agcaaccgtt tgacgtgatc gtggcgtacc gggtagatcg gttgacccga
     1081 tcgatccggc atcttcagca gctggtccac tgggccgagg accacaagaa gctggtcgtc
     1141 tccgcgaccg aagcgcactt cgatacgacg acgccgtttg cggcggtcgt catcgcgctt
     1201 atgggaacgg tggcgcagat ggaattagaa gcgatcaaag agcggaaccg ttcggctgcg
     1261 catttcaata tccgcgccgg gaaataccga ggatccctgc cgccgtgggg atacctgcct
     1321 acgcgcgtgg acggggagtg gcggctggtg ccggaccctg tgcagcgaga gcgcatcctc
     1381 gaggtgtatc accgcgtcgt cgacaaccac gagccgctgc atctggtggc ccacgacctg
     1441 aaccggcgtg gtgtcctgtc gccgaaggac tacttcgcgc agctgcaagg ccgcgagccg
     1501 cagggccggg agtggtcggc taccgcgctg aagcgatcga tgatctccgg ggcgatgctc
     1561 gggtacgcga ctctgaacgg taagaccgtc cgagacgacg acggagcccc gctggtgcgg
     1621 gctgagccga tcctgacccg tgagcagctg gaggcgctgc gcgccgagct cgtgaagacc
     1681 tcccgggcga agcccgcggt gtctaccccg tcgctgctgc tgcgggtgtt gttctgcgcg
     1741 gtgtgcgggg agcccgcgta caagttcgcc gggggaggac gtaagcaccc gcgctaccgc
     1801 tgccgctcga tggggttccc gaagcactgc gggaacggca cggtggcgat ggccgagtgg
     1861 gacgcgttct gcgaggagca ggtactggat ctgctcgggg acgcggagcg tctggagaaa
     1921 gtctgggtag cgggctcgga ctccgcggtc gaactcgcgg aggtgaacgc ggagctggtg
     1981 gacctgacgt cgctgatcgg ctccccggcc taccgggcgg gctctccgca gcgagaagca
     2041 ctggatgccc gtattgcggc gctggccgcg cggcaagagg agctggaggg cctggaggct
     2101 cgcccgtctg gctgggagtg gcgcgaaacc gggcagcggt tcggggactg gtggcgggag
     2161 caggacaccg cggcaaagaa cacctggctt cggtcgatga acgttcggct gacgttcgac
     2221 gtccgcggcg ggctgactcg cacgatcgac ttcggggatc ttcaggagta cgagcagcat
     2281 ctcaggctcg gcagcgtggt cgaacggcta cacaccggga tgtcgtaata agcttatctc
     2341 gtctctgtta tgcttaagaa gttcaatgtt tcgtttcatg taaaactttg gtggtttgtg
     2401 ttttggggcc ttgtataatc cctgatgaat aagtgttcta ctatgtttcc gttcctgtta
     2461 tctctttctt tctaatgaca agtcgaactt cttctttatc atcgcttcgt ttttattatc
     2521 tgtgcttctt ttgtttaata cgcctgcaaa gtgactcgac tctgtttagt gcagttctgc
     2581 gaaacttgta aatagtccaa ttgttggcct ctagtaatag atgtagcgaa agtgttgagc
     2641 tgttgggttc taaggatggc ttgaacatgt taatctttta ggttctgagt atgatgaaca
     2701 ttcgttgttg ctaagaaatg cctgtaatgt cccacaaatg tagaaaatgg ttcgtacctt
     2761 tgtccaagca ttgatatgtc tgatgagagg aaactgcaag atactgagct tggtttaacg
     2821 aaggagaggc agtttcttcc ttccaaagca tttcatttga caatgccttg atcatcttaa
     2881 gtagagtttc tgttgtggaa agtttgaaac tttgaagaaa cgactctcaa gtaaattgat
     2941 gatcacaagt gaaagtgtat gttcgcttga gacctccgat cccctgaatt ctgcagatat
     3001 ccatcacact ggcggccgct cgagcatgca tctagagggc ccaattcgcc ctatagtgag
     3061 tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     3121 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     3181 gcccgcaccg atcgcccttc ccaacagttg cgcagcctat acgtacggca gtttaaggtt
     3241 tacacctata aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt
     3301 gacacgccgg ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa
     3361 gtctcccgtg aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc
     3421 accgatatgg ccagtgtgcc ggtctccgtt atcggggaag aagtggctga tctcagccac
     3481 cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc
     3541 atgagattat caaaaaggat cttcacctag atccttttca cgtagaaagc cagtccgcag
     3601 aaacggtgct gaccccggat gaatgtcagc tactgggcta tctggacaag ggaaaacgca
     3661 agcgcaaaga gaaagcaggt agcttgcagt gggcttacat ggcgatagct agactgggcg
     3721 gttttatgga cagcaagcga accggaattg ccagctgggg cgccctctgg taaggttggg
     3781 aagccctgca aagtaaactg gatggctttc ttgccgccaa ggatctgatg gcgcagggga
     3841 tcaagctctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     3901 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3961 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4021 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     4081 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4141 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4201 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4261 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4321 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4381 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     4441 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4501 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4561 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4621 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgaattatt
     4681 aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4741 cgcatcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4801 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4861 gcacgtgagg agggccacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg
     4921 cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt
     4981 ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca
     5041 ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct
     5101 gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat
     5161 gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa
     5221 ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctaaaacttc atttttaatt
     5281 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5341 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5401 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5461 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5521 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5581 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5641 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5701 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5761 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5821 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5881 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5941 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6001 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6061 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     6121 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     6181 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     6241 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     6301 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     6361 tcacacagga aacagctatg accatgatta cgccaagcta tttaggtgac gcgttagaat
     6421 actcaagcta tgcatcaagc ttggtaccga gctcggatcc actagtaacg gccgccagtg
     6481 tgctggaatt cagg
//
