LOCUS       Exported                5257 bp DNA     circular SYN 10-SEP-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5257)
  AUTHORS   11111111
  TITLE     Direct Submission
  JOURNAL   Exported Sep 10, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5257
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        256..274
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    275..299
                     /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)."
     CDS             349..366
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             385..399
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             478..522
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     terminator      574..621
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      658..1113
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(1228..2088)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2089..2181)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2262..2850
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3036..3175
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3277..3468)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4040..5122)
                     /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"
     promoter        complement(5123..5200)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca
       61 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
      121 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
      181 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatcgatc
      241 tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
      301 ctctagaaat aattttgttt aactttaaga aggagatata ccatggcaca tcaccaccac
      361 catcacgtgg gtaccggttc gaatgatgac gacgacaaga gtccggatcc caattgggag
      421 ctcgtgtaca cggcgcgcct gcaggtcgac aagcttgcgg ccgcactcga gtctggtaaa
      481 gaaaccgctg ctgcgaaatt tgaacgccag cacatggact cgtctactag cgcagcttaa
      541 ttaacctagg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
      601 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg
      661 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
      721 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
      781 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
      841 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
      901 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
      961 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     1021 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     1081 cgaattttaa caaaatatta acgtttacaa tttctggcgg cacgatggca tgagattatc
     1141 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1201 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     1261 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     1321 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     1381 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     1441 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     1501 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     1561 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     1621 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     1681 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     1741 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     1801 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     1861 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     1921 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     1981 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     2041 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     2101 tcaatcatga ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2161 gtatttagaa aaataaacaa ataggtcatg accaaaatcc cttaacgtga gttttcgttc
     2221 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2281 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2341 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2401 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2461 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2521 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2581 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2641 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2701 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2761 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2821 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2881 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     2941 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3001 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     3061 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
     3121 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     3181 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     3241 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     3301 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     3361 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     3421 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     3481 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     3541 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgatg gtaaacaact
     3601 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     3661 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     3721 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     3781 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     3841 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     3901 cctcaacgac aggagcacga tcatgctagt catgccccgc gcccaccgga aggagctgac
     3961 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     4021 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4081 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     4141 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     4201 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     4261 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     4321 tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     4381 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     4441 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     4501 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     4561 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     4621 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     4681 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     4741 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     4801 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     4861 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     4921 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     4981 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     5041 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     5101 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     5161 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     5221 ttatgcgact cctgcattag gaagcagccc agtagta
//
