LOCUS       sequence-266747-        6418 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 6418)
  AUTHORS   LaRonde-LeBlanc NA, Wolberger C
  TITLE     Structure of HoxA9 and Pbx1 bound to DNA: Hox hexapeptide and DNA 
            recognition anterior to posterior.
  JOURNAL   Genes Dev 2003 Aug 15;17(16):2060-72.
  PUBMED    12923056
REFERENCE   2  (bases 1 to 6418)
  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: "Genes Dev";
            date: "2003-08-15"; volume: "17(16)"; pages: "2060-72"
COMMENT     Sequence Label: pPBX1-318
FEATURES             Location/Qualifiers
     source          1..6418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1150..1167
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1220..1238)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1234..1281
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1318..1773
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1405..1424)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1615..1636
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1799..1903
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1904..1972,1973..2764)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2122..2141)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2935..3523
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3424..3443
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3677..3694
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3709..3848
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3774..3793)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3934..3956
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3950..4141)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4950..6032)
                     /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     5994..6013
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6033..6110)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6316..6335
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggacgagcag cccaggctga tgcattccca
      121 tgctggggtc gggatggccg gacaccccgg cctgtcccag cacttgcagg atggggccgg
      181 agggaccgag ggggagggcg ggaggaagca ggacattgga gacattttac agcaaattat
      241 gaccatcaca gaccagagtt tggatgaggc gcaggccaga aaacatgctt taaactgcca
      301 cagaatgaag cctgccttgt ttaatgtgtt gtgtgaaatc aaagaaaaaa cagttttgag
      361 tatccgagga gcccaggagg aggaacccac agacccccag ctgatgcggc tggacaacat
      421 gctgttagcg gaaggcgtgg cggggcctga gaagggcgga gggtcggcgg cagcggcggc
      481 agcggcggcg gcttctggag gggcaggttc agacaactca gtggagcatt cagattacag
      541 agccaaactc tcacagatca gacaaatcta ccatacggag ctggagaaat acgagcaggc
      601 ctgcaacgag ttcaccaccc acgtgatgaa tctcctgcga gagcaaagcc ggaccaggcc
      661 catctcccca aaggagattg agcggatggt cagcatcatc caccgcaagt tcagctccat
      721 ccagatgcag ctcaagcaga gcacgtgcga ggcggtgatg atcctgcgtt cccgatttct
      781 ggatgcgcgg cggaagagac ggaatttcaa caagcaagcg acagaaatcc tgaatgaata
      841 tttctattcc catctcagca acccttaccc cagtgaggaa gccaaagagg agttagccaa
      901 gaagtgtggc atcacagtct cccaggtatc aaactggttt ggaaataagc gaatccggta
      961 caagaagaac ataggtaaat ttcaagagga agccaatatt tatgctgcca aaacagctgt
     1021 cactgctacc aatgtgtcag cccatggaag ccaagctaac tcgccctcaa ctcccaactc
     1081 ggctggtgga tacccttcgc catgttatca gccagacagg aggatacagt gacggatccg
     1141 cagccagtgc accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa
     1201 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     1261 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg
     1321 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1381 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1441 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1501 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     1561 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1621 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     1681 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     1741 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     1801 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1861 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     1921 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     1981 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     2041 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     2101 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     2161 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     2221 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     2281 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     2341 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2401 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     2461 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     2521 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     2581 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     2641 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     2701 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     2761 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     2821 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2881 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2941 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3001 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3061 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     3121 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3181 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3241 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3301 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3361 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3421 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3481 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3541 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3601 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3661 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     3721 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
     3781 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     3841 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     3901 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     3961 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     4021 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     4081 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     4141 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
     4201 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
     4261 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
     4321 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
     4381 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
     4441 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
     4501 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
     4561 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc
     4621 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     4681 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     4741 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     4801 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     4861 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     4921 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4981 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5041 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     5101 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     5161 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     5221 actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     5281 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     5341 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     5401 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     5461 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     5521 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     5581 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     5641 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     5701 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     5761 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     5821 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     5881 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     5941 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     6001 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     6061 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     6121 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     6181 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     6241 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     6301 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     6361 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaat
//
