LOCUS       Exported                6238 bp ds-DNA     circular SYN 03-FEB-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6238)
  AUTHORS   Keoni Gandall
  TITLE     Direct Submission
  JOURNAL   
FEATURES             Location/Qualifiers
     source          1..6238
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     16..32
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      33..64
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      complement(163..751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(942..1802)
                     /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(1803..1907)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1972..2019
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2024..2040)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2199..2449
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(2452..3207)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      3298..3517
                     /label=oriS
                     /note="secondary origin of replication for the bacterial F 
                     plasmid"
     misc_feature    3872..4046
                     /label=lacOP
     protein_bind    3937..3958
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3973..4003
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4011..4027
                     /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)."
     primer_bind     4035..4051
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4124..4969
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    5077..5228
                     /label=nnpp promoters
     CDS             5268..6083
                     /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"
     terminator      6193..6236
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 cacctgcacc agtcagtaaa acgacggcca gtagtcaaaa gcctccgacc ggaggctttt
       61 gacttggttc aggtggagtg ggagtagtct tcgccatcgc cttggcaggt gatgacacac
      121 attaacaaat ttcgtgagga gtctccagaa gaatgccatt aatttccata ggctccgccc
      181 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
      241 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
      301 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
      361 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
      421 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
      481 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
      541 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
      601 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
      661 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
      721 gcagattacg cgcagaaaaa aaggatctca agaaggccta ctattagcaa caacgatcct
      781 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      841 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      901 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      961 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1021 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1081 cgagaaccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1141 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1201 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1261 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1321 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1381 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1441 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1501 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1561 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1621 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1681 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1741 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1801 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1861 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac ctgcaccagc
     1921 gatgttgaag accacgctga ggtgtcaatc gtcggagccg ctgagcaata actagcataa
     1981 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc atggtcatag ctgtttcctg
     2041 agagcttggc aggtgtcgcg tatgcatcga tcacgtgtcc acgttcttta atagtggact
     2101 cttgttccaa actggaacaa cactcggatc gatccggcgc gcaccgtggg aaaaactcca
     2161 ggtagaggta cacacgcgga tagccaattc agagtaataa actgtgataa tcaaccctca
     2221 tcaatgatga cgaactaacc cccgatatca ggtcacatga cgaagggaaa gagaaggaaa
     2281 tcaactgtga caaactgccc tcaaatttgg cttccttaaa aattacagtt caaaaagtat
     2341 gagaaaatcc atgcaggctg aaggaaacag caaaactgtg acaaattacc ctcagtaggt
     2401 cagaacaaat gtgacgaacc accctcaaat ctgtgacaga taaccctcag actatcctgt
     2461 cgtcatggaa gtgatatcgc ggaaggaaaa tacgatatga gtcgtctggc ggcctttctt
     2521 tttctcaatg tatgagaggc gcattggagt tctgctgttg atctcattaa cacagacttg
     2581 caggaagcgg cggcggaagt caggcatacg ctggtaactt tgaggcagct ggtaacgctc
     2641 tatgatccag tcgattttca gagacacgat gcctgagcca tccggcttac gatactgaca
     2701 cagggattcg tataaacgca tggcatacgg attggtgatt tcttttgttt cactaagccg
     2761 aaactgcgta aaccggttct gtaacccgat aaagaaggga atgagatatg ggttgatatg
     2821 tacactgtaa agccctctgg atggactgtg cgcacgtttg ataaaccaag gaaaagattc
     2881 atagcctttt tcatcaccgg catcctcttc agggcgataa aaaaccactt ccttccccgc
     2941 gaaactcttc aatgcctgcc gtatatcctt actggcttcc gcagaggtca atccgaatat
     3001 ttcagcatat ttagcaacat ggatctcgca gataccgtca tgttcctgta gggtgccatc
     3061 agattttctg atctggtcaa cgaacagata cagcatacgt ttttgatccc gggagagact
     3121 atatgccgcc tcagtgaggt cgtttgactg gacgattcgc gggctatttt tacgtttctt
     3181 gtgattgata accgctgttt ccgccatgac agatccatgt gaagtgtgac aagtttttag
     3241 attgtcacac taaataaaaa agagtcaata agcagggata actttgtgaa aaaacagctt
     3301 cttctgaggg caatttgtca cagggttaag ggcaatttgt cacagacagg actgtcattt
     3361 gagggtgatt tgtcacactg aaagggcaat ttgtcacaac accttctgta gaaccagcat
     3421 ggataaaggc ctacaaggcg ctctaaaaaa gaagatctaa aaactataaa aaaaataatt
     3481 ataaaaatat ccccgtggat aagtggataa ccccaaggga agttttttca ggcatcgtgt
     3541 gtaagcagaa tatataagtg ctgttccctg gtgcttcctc gctcactcga aattcccggg
     3601 gatagcttta tgcttgtaaa ccgttttgtg aaaaaatttt taaaataaaa aaggggacct
     3661 ctagggtccc caattaatta gtaatataat ctattaaagg tcattcaaaa ggtcatccac
     3721 cggatcaatt cccctgctcg cgcaggctgg gtgccaagct ctcgggtaac atcaaggccc
     3781 gatccttgga gcccttgccc tcccgcacga tgatcgtgcc gtgatcgaaa tccagatcct
     3841 tgacccgcag ttgcaaaccc tcactgatcc gattcattaa tgcagctggc acgacaggtt
     3901 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     3961 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     4021 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg gtacctgtcg
     4081 cggcaacgcg ctaacagacg tagtaagaac caccagcatt gtaatgctgg ctaaagtcac
     4141 tttcctgagc tgtataacga tgagcgattt tactttttct ggctatgaat tggcctgctt
     4201 tgtaacacac tccggtctat cccgtagcgc cgggcatatc ctgtcgcaat gtgcaaatct
     4261 cgcggcaaca accagtgaat acttcattca caagcctcac cgcctgatcg cggcagaaac
     4321 tggttatagc caatcaaccg tcgttcgtgc attccgtgaa gctgtaaaca aaggtatcct
     4381 gtctgtagag attgttatcg gcgatcaccg tgaacgtcgc gctaacctgt accggtttac
     4441 accatccttt ttggccttcg cacaacaagc caaaaatgcg ctgatagaaa gcaaattaaa
     4501 gatctcttca gcggcaacca aggttaaagc tgttctcgct aagacattgg ctttatttaa
     4561 ttttttatcc acacccccat gtcaaaatga taccccctcc ccctgtcagg atgacgtggc
     4621 aataaagaat aagaagtcac aagttaaaaa aacaaaaaga tcagtttccg gcggtgccgg
     4681 aacaaccagc ctcaaaaaat tgacttcatg gatcgctaag gcaaaagcaa aggctgacaa
     4741 tctgcggtta tccaaaaaac gcactcaaaa acatgagttc aagcagaaag tagaggcggc
     4801 tgcgcggaaa tatgcttacc tgaagaacaa gcgttcgcct gatattggcg ggatatcaaa
     4861 cttcgataac ctaccgcatt gcatgacggt aaacgaagct cttaatgcgg ttttagccaa
     4921 aaataaagat aacgaacaat ggggtatacc ggcaggattc agagggtaat gaattgctct
     4981 aattataacc atgcatactt tcaacacctc tagtttgcca tgaggcaaac tcataggtgt
     5041 cctggtaaga ggacactgtt gccaaaactg gacgccccat tattgcaatt aataaacaac
     5101 taacggacaa ttctacctaa caataagtgg agttgcggcc ggccgcgatc cgctagcaaa
     5161 gccacgttgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     5221 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     5281 aacgggaaac gtcttgctcg cgcccgcgat taaattccaa catggatgct gatttatatg
     5341 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg
     5401 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     5461 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     5521 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccgggaaaa
     5581 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     5641 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     5701 gcgtatttcg cctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     5761 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac
     5821 tgttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     5881 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     5941 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     6001 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     6061 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac tggcgagctc
     6121 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     6181 tcacgaggca gaatttcaga taaaaaaaat ccttagcttt cgctaaggat gatttctg
//
