LOCUS       Exported                4629 bp DNA     circular SYN 21-JUL-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4629)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4629
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    95..116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        131..161
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    169..185
                     /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             205..570
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWKL"
     promoter        230..248
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    261..368
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     285..301
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(335..351)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(377..395)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        668..847
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             881..1675
                     /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      1715..1967
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    1991..2013
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     rep_origin      complement(2080..2668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2769..3584)
                     /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"
     rep_origin      3875..4310
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    4461..4485
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
       61 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      121 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      181 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
      241 actaaaggga acaaaagctg gagctccacc gcggtggcgg ccgctctaga actagtggat
      301 cccccgggct gcaggaattc gatatcaagc ttatcgatac cgtcgacctc gagggggggc
      361 ccggtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
      421 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
      481 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
      541 cgcagcctga atggcgaatg gaaattgtaa gcgttaatgt tatcgatcat gagcggagaa
      601 ttaagggagt cacgttatga cccccgccga tgacgcggga caagccgttt tacgtttgga
      661 actgacagaa ccgcaacgtt gaaggagcca ctgagccgcg ggtttctgga gtttaatgag
      721 ctaagcacat acgtcagaaa ccattattgc gcgttcaaaa gtcgcctaag gtcactatca
      781 gctagcaaat atttcttgtc aaaaatgctc cactgacgtt ccataaattc ccctcggtat
      841 ccaattagag tctcatattc actctcaact cgatcgaggc atgattgaac aagatggatt
      901 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
      961 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     1021 ttttgtcaag accgacctgt ccggtgccct gaatgaactc caagacgagg cagcgcggct
     1081 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     1141 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     1201 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     1261 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     1321 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     1381 agccgaactg ttcgccaggc tcaaggcgcg gatgcccgac ggcgaggatc tcgtcgtgac
     1441 ccacggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     1501 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     1561 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     1621 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     1681 actctggggt tcggactcta gctagagtca agcagatcgt tcaaacattt ggcaataaag
     1741 tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt atcatataat ttctgttgaa
     1801 ttacgttaag catgtaataa ttaacatgta atgcatgacg ttatttatga gatgggtttt
     1861 tatgattaga gtcccgcaat tatacattta atacgcgata gaaaacaaaa tatagcgcgc
     1921 aaactaggat aaattatcgc gcgcggtgtc atctatgtta ctagatcgac cggcatgcaa
     1981 gctgataacg ttacaccaca atatatcctg ccaagatctc atgtgagcaa aaggccagca
     2041 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2101 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2161 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2221 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2281 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2341 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2401 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2461 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2521 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2581 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2641 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2701 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atggttacaa ccaattaacc
     2761 aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     2821 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     2881 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     2941 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     3001 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca
     3061 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     3121 cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca
     3181 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     3241 tcaggatatt cttctaatac ctggaatgct gttttccctg ggatcgcagt ggtgagtaac
     3301 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     3361 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     3421 ttcagaaaca actctggcgc atcgggcttc ccatacaatc ggtagattgt cgcacctgat
     3481 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     3541 aatcgcggcc ttgagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta
     3601 ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg
     3661 taacatcaga gattttgagc cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag
     3721 ttgaaggatc agatcacgca tcttcccgac aacgcagacc gttccgtggc aaagcaaaag
     3781 ttcaaaatca ccaactggtc cacctacaac aaagctctca tcaaccgtgg ctccctcact
     3841 ttctggctgg atgatggggc gattcaggcg atccccatcc aacagcccgc cgtcgagcgg
     3901 gcttttttat ccccggaagc ctgtggatag agggtagtta tccacgtgaa accgctaatg
     3961 ccccgcaaag ccttgattca cggggctttc cggcccgctc caaaaactat ccacgtgaaa
     4021 tcgctaatca gggtacgtga aatcgctaat cggagtacgt gaaatcgcta ataaggtcac
     4081 gtgaaatcgc taatcaaaaa ggcacgtgag aacgctaata gccctttcag atcaacagct
     4141 tgcaaacacc cctcgctccg gcaagtagtt acagcaagta gtatgttcaa ttagcttttc
     4201 aattatgaat atatatatca attattggtc gcccttggct tgtggacaat gcgctacgcg
     4261 caccggctcc gcccgtggac aaccgcaagc ggttgcccac cgtcgagcgc ctttgcccac
     4321 aacccggcgg ccggccgcaa cagatcgttt tataaatttt tttttttgaa aaagaaaaag
     4381 cccgaaaggc ggcaacctct cgggcttctg gatttccgat ccccggaatt agatctcaaa
     4441 caaacacata cagcgactta gtttacccgc caatatatcc tgtcaaggcc ttcatgttct
     4501 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4561 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4621 gcccaatac
//
