LOCUS       Exported                5000 bp DNA     circular SYN 05-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5000)
  AUTHORS   11111111
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1..2346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          4781..5000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             complement(11..33)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(48..72)
                     /label=lac operator
                     /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)."
     promoter        complement(73..91)
                     /label=T7 promoter promoter for bacteriophage T7 RNA p
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        400..477
                     /label=lacI promoter
     CDS             601..969
                     /codon_start=1
                     /gene="lacI"
                     /product="Lac repressor"
                     /label=lacI
                     /protein_id="AVN68413.1"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSR
                     ADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPA
                     LFLDVSDQTPINSIIFLP"
     gene            601..969
                     /gene="lacI"
                     /label=lacI
     protein_bind    1574..1595
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    complement(1946..2314)
                     /label=plasmid partitioning sequence - par
                     /note="plasmid partitioning sequence - par"
     primer_bind     complement(2356..2372)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2380..2396
                     /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)."
     promoter        complement(2404..2434)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2449..2470
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2758..3346)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3630..4424)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     terminator      complement(4819..4866)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4983..5000)
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
ORIGIN
        1 catatgtata tctccttctt aaagttaaac aaaattattt ctagagggga attgttatcc
       61 gctcacaatt cccctatagt gagtcgtatt aatttcgcgg gatcgagatc tcgatcctct
      121 acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata
      181 tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg agcgcttgtt
      241 tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc atctccttgc
      301 atgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc
      361 taatgcagga gtcgcataag ggagagcgtc gagatcccgg acaccatcga atggcgcaaa
      421 acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg
      481 aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc
      541 cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg
      601 atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg
      661 ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg
      721 gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga
      781 agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg
      841 ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact
      901 aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttt
      961 ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca
     1021 aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg
     1081 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag
     1141 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc
     1201 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg
     1261 gctgcgcgtt ggtgcggaca tctcggtagt gggatacgac gataccgaag acagctcatg
     1321 ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt
     1381 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt
     1441 ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc
     1501 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     1561 agcgcaacgc aattaatgta agttagctca ctcattaggc accgggatct cgaccgatgc
     1621 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     1681 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     1741 gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg
     1801 cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac
     1861 gtttcggcga gaagcaggcc attatcgccg gcatggcggc cccacgggtg cgcatgatcg
     1921 tgctcctgtc gttgaggacc gatcacgggc aaatcgctga atattccttt tgtctccgac
     1981 catcaggcac ctgagtcgct gtctttttcg tgacattcag ttcgctgcgc tcacggctct
     2041 ggcagtgaat gggggtaaat ggcactacag gcgcctttta tggattcatg caaggaaact
     2101 acccataata caagaaaagc ccgtcacggg cttctcaggg cgttttatgg cgggtctgct
     2161 atgtggtgct atctgacttt ttgctgttca gcagttcctg ccctctgatt ttccagtctg
     2221 accacttcgg attatcccgt gacaggtcat tcagactggc taatgcaccc agtaaggcag
     2281 cggtatcatc aacaggctta cccgtcttac tgtcgaattc aatcgaattc ccgcggccag
     2341 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     2401 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2461 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2521 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2581 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2641 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2701 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2761 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2821 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2881 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2941 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3001 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3061 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3121 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3181 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3241 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3301 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3361 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3421 gagattatca aaaaggatct tcacctagat ccttttgggg tgggcgaaga actccagcat
     3481 gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa
     3541 cctttcatag aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg
     3601 gtcggtcatt tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag
     3661 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
     3721 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc
     3781 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
     3841 ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catccgcgcc
     3901 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc
     3961 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg
     4021 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
     4081 atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg
     4141 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga
     4201 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cttggagttc attcagggca
     4261 ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg
     4321 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
     4381 caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat
     4441 cctgtctctt gatcagatct tgatcccctg cgccatcaga tccttggcgg caagaaagcc
     4501 atccagttta ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc
     4561 ggttcgcttg ctgtccataa aaccgcccag tctagctatc gccatgtaag cccactgcaa
     4621 gctacctgct ttctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca
     4681 ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtgttccg cttcctttag
     4741 cagcccttgc gccctgagtg cttgcggcag cgtgaagcta gtgattctcg acgatccgga
     4801 tatagttcct cctttcagca aaaaacccct caagacccgt ttagaggccc caaggggtta
     4861 tgctagttat tgctcagcgg tggcagcagc caactcagct tcctttcggg ctttgttagc
     4921 agccggatca agcttcgaat tccatggtac cagctgcaga tctcgaggtc ggatccctcg
     4981 aggtgatggt gatggtgatg
//
