LOCUS       sequence-217374-        6104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ty3 Gag3-His tagged, codon optimized for expression in E. coli; . 
            IPTG inducible.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6104)
  TITLE     Ty3 capsid codon optimized for E. coli expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6104)
  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            "Unpublished"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: pNB3033
FEATURES             Location/Qualifiers
     source          1..6104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             878..895
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(948..966)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      962..1009
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1046..1501
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1133..1152)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1343..1364
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1593..2408)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1593..2408)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2316..2335
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2400..2419
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2530..3118
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3019..3038
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3272..3289
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3304..3443
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3369..3388)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3529..3551
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3545..3736)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4545..5627)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5589..5608
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5628..5705)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5911..5930
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6018..6037
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6018..6036
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6037..6061
                     /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)."
     RBS             6076..6098
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 tatgtcattc atggaccaga ttccgggcgg gggtaactat cctaaattgc cagtagaatg
       61 tttgccgaat tttcccattc aaccaagtct gacctttcgc gggagaaacg atagccacaa
      121 actgaagaat ttcattagcg agattatgct caacatgtcg atgatctctt ggcctaacga
      181 tgcgtctaga attgtgtact gccgtcgtca tttacttaat ccagctgctc agtgggctaa
      241 tgactttgtg caagaacagg gtattctcga gattacgttc gatacattta tccaggggct
      301 gtatcaacac ttttataaac cgcctgatat caataaaatc tttaacgcca tcacgcagct
      361 gtccgaggca aaattaggca ttgaacgtct gaatcaacgg tttcggaaaa tttgggatcg
      421 catgccacca gatttcatga cagaaaaggc cgcaattatg acgtataccc ggttactgac
      481 gaaagagacc tataatattg tacgtatgca taagccggag accctgaaag atgcgatgga
      541 ggaagcctac cagaccactg cccttaccga acgattcttc cctggctttg aactggacgc
      601 ggacggagat accatcatag gcgcaaccac tcacttgcag gaagagtacg attctgatta
      661 cgattcggaa gataacctga cccaaaatgg ctacgttcac actgttagga cccgtcgttc
      721 atacaataag cccatgagca accatcggaa ccgcagaaac aacaacccga gccgagaaga
      781 atgcatcaaa aaccgtctgt gcttttattg taagaaagaa ggccatcgac tgaatgaatg
      841 ccgtgcccgt aaagcaagct ctaaccgtag cctcgagcac caccaccacc accactgaga
      901 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
      961 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     1021 aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1081 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1141 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     1201 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     1261 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     1321 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     1381 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     1441 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     1501 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1561 attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga
     1621 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
     1681 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
     1741 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
     1801 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta
     1861 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     1921 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg
     1981 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     2041 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc
     2101 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     2161 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     2221 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     2281 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     2341 aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata
     2401 tggctcataa caccccttgt attactgttt atgtaagcag acagttttat tgttcatgac
     2461 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2521 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2581 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2641 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     2701 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2761 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2821 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2881 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2941 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3001 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3061 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3121 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     3181 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     3241 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     3301 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc
     3361 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     3421 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     3481 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     3541 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     3601 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     3661 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     3721 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
     3781 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
     3841 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
     3901 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
     3961 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
     4021 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
     4081 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag
     4141 taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc
     4201 gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac
     4261 cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga
     4321 tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca
     4381 cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc
     4441 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc
     4501 ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc
     4561 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4621 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
     4681 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
     4741 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
     4801 gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg
     4861 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
     4921 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
     4981 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
     5041 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
     5101 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
     5161 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
     5221 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
     5281 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
     5341 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
     5401 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
     5461 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
     5521 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
     5581 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     5641 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
     5701 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag
     5761 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc
     5821 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     5881 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     5941 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatcga
     6001 tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc
     6061 ccctctagaa ataattttgt ttaactttaa gaaggagata taca
//
