LOCUS       sequence-213926-        6108 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 6108)
  TITLE     Ty3 capsid codon optimized for E. coli expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6108)
  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            "Unpublished"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..6108
                     /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(1594..2409)
                     /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(1594..2409)
                     /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     2317..2336
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2401..2420
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2531..3119
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3020..3039
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3273..3290
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3305..3447
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3373..3392)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3533..3555
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3549..3740)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4549..5631)
                     /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     5593..5612
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5632..5709)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5915..5934
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6022..6041
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6022..6040
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6041..6065
                     /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             6080..6102
                     /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 gtttacaatt
     1501 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1561 cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg
     1621 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     1681 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     1741 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     1801 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
     1861 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     1921 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     1981 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     2041 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     2101 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     2161 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     2221 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     2281 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     2341 taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat
     2401 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     2461 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2521 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2581 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2641 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     2701 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2761 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     2821 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     2881 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     2941 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3001 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3061 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3121 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3181 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3241 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3301 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatatg
     3361 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     3421 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     3481 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     3541 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     3601 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     3661 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     3721 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta
     3781 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc
     3841 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga
     3901 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt
     3961 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
     4021 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
     4081 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
     4141 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc
     4201 acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg
     4261 ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg
     4321 ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc
     4381 ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc
     4441 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     4501 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     4561 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4621 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     4681 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     4741 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     4801 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt
     4861 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     4921 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     4981 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     5041 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     5101 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     5161 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     5221 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     5281 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     5341 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     5401 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     5461 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     5521 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     5581 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     5641 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     5701 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     5761 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     5821 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     5881 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     5941 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     6001 tcgatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca
     6061 attcccctct agaaataatt ttgtttaact ttaagaagga gatataca
//
