LOCUS       sequence-208949-        6179 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6179)
  TITLE     HOX protein coding clones not referenced in publications
REFERENCE   2  (bases 1 to 6179)
  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: HOXD9 (human)  pET
FEATURES             Location/Qualifiers
     source          1..6179
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     209..228
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    228..252
                     /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             267..289
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             308..325
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             335..352
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             356..388
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             1252..1269
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1322..1340)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1336..1383
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1420..1875
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1507..1526)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1717..1738
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1967..2782)
                     /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(1967..2782)
                     /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     2690..2709
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2774..2793
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2904..3492
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3393..3412
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3646..3663
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3678..3817
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3743..3762)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3903..3925
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3919..4110)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4919..6001)
                     /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     5963..5982
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6002..6079)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
       61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
      181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
      241 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat ataccatggg
      301 cagcagccat catcatcatc atcacagcag cggcctggtg ccgcgcggca gccatatggc
      361 tagcatgact ggtggacagc aaatgggtcg cggatccatg tcttccagtg gcaccctcag
      421 caactactac gtggactcgc ttataggcca tgagggcgac gaggtgttcg cggcgcgctt
      481 cgggccgccg gggccaggcg cgcagggccg gcctgcaggt gtggctgatg gcccggccgc
      541 caccgccgcc gagttcgcct cgtgtagttt tgcccccaga tcggccgtgt tctctgcctc
      601 gtggtccgcg gtgccctccc agcccccggc agcggcggcg atgagcggcc tctaccaccc
      661 gtacgttccc ccgccgcccc tggccgcctc tgcctccgag cccggccgct acgtgcgctc
      721 ctggatggag ccgctgcccg gcttcccggg cggtgcgggc ggtggcggtg gtggtggagg
      781 cggcggtccg ggccgcggtc ccagccctgg ccccagcggc ccagccaacg ggcgccacta
      841 cgggattaag cctgaaaccc gagcggcccc ggcccccgcc acggccgcct ccaccacctc
      901 ctcctcctcc acttccttat cctcctcctc caaacggact gagtgctccg tggcccggga
      961 gtcccagggg agcagcggcc ccgagttctc gtgcaactcg ttcctgcagg agaaggcggc
     1021 agcggcgacg gggggaaccg ggcctggggc agggatcggg gccgcgactg ggacgggcgg
     1081 ctcgtcggag ccctcagctt gcagcgacca cccgatccca ggctgttcgc tgaaggagga
     1141 ggagaagcag cattcgcagc cgcagcagca gcagcaactt gacccaaaca accccgccgc
     1201 gaactggatc cgaattcgag ctccgtcgac aagcttgcgg ccgcactcga gcaccaccac
     1261 caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     1321 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1381 ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat
     1441 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     1501 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     1561 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     1621 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     1681 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     1741 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     1801 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     1861 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     1921 atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat
     1981 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     2041 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     2101 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     2161 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     2221 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     2281 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     2341 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     2401 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     2461 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     2521 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     2581 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     2641 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     2701 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg
     2761 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     2821 ttattgttca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     2881 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2941 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3001 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     3061 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3121 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3181 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3241 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3301 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3361 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3421 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3481 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3541 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3601 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3661 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3721 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     3781 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     3841 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3901 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     3961 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     4021 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     4081 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     4141 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     4201 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     4261 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
     4321 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
     4381 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     4441 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     4501 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     4561 gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     4621 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     4681 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     4741 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     4801 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     4861 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     4921 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4981 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     5041 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     5101 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     5161 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc
     5221 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     5281 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     5341 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     5401 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     5461 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     5521 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     5581 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     5641 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     5701 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     5761 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     5821 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     5881 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     5941 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     6001 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     6061 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     6121 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatg
//
