LOCUS       Exported                6224 bp ds-DNA     circular SYN 08-MAR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6224)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 11, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6224
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             129..989
                     /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"
     primer_bind     complement(347..366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1649..1668
                     /label=oJDW 1410
     primer_bind     1921..1938
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1944..1961
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1945..1961
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   1987..2049
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     2227..2260
                     /label=oJDW 1372 attR4 + SpeI
     protein_bind    2251..2375
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2416..2721)
                     /label=ccdb
     misc_feature    complement(2495..2515)
                     /label=RFA Seq Primer 2
     primer_bind     complement(2495..2515)
                     /label=oJDW 93
     CDS             complement(3063..3721)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGR"
     primer_bind     3636..3655
                     /label=oJDW 253
     misc_feature    complement(3773..3793)
                     /label=RFA Seq Primer 1
     promoter        complement(3775..3805)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(3831..3954)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3962..3978)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(3962..3978)
                     /label=M13 For -20
     primer_bind     complement(3962..3978)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3969..3990)
                     
     primer_bind     complement(3987..4003)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3988..4004)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        4024..4340
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4191..4326
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4253..4272
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4388..4435
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4454..5248
                     /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"
     primer_bind     complement(4508..4527)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5118..5137
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5288..5307)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5332..5380
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(5596..5617)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     repeat_region   5901..5935
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     6115..6134
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgct gtacagctta agtcatgttc tttcctgcgt tatcccctga ttctgtggat
     1861 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     1921 agcgagtcag tgagcgagga agctgtaaaa cgacggccag tagctccacc gccgcagcta
     1981 gattaaccct agaaagataa tcatattgtg acgtacgtta aagataatca tgcgtaaaat
     2041 tgacgcatgt gttttatcgg tctgtatatc gaggtttatt tattaatttg aatagatatt
     2101 aagttttatt atatttacac ttacatacta ataataaatt caacaaacaa tttatttatg
     2161 tttatttatt tattaaaaaa aaacaaaaac tcaaaatttc ttctataaag taacaaaact
     2221 tttatgacta gtcatgatta cgccaagcta tcaactttgt atagaaaagt tgaacgagaa
     2281 acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa acagactaca
     2341 taatactgta aaacacaaca tatccagtca ctatggtcga cctgcagact ggctgtgtat
     2401 aagggagcct gacatttata ttccccagaa catcaggtta atggcgtttt tgatgtcatt
     2461 ttcgcggtgg ctgagatcag ccacttcttc cccgataacg gagaccggca cactggccat
     2521 atcggtggtc atcatgcgcc agctttcatc cccgatatgc accaccgggt aaagttcacg
     2581 ggggacttta tctgacagca gacgtgcact ggccaggggg atcaccatcc gtcgcccggg
     2641 cgtgtcaata atatcactct gtacatccac aaacagacga taacggctct ctcttttata
     2701 ggtgtaaacc ttaaactgca tttcaccagc ccctgttctc gtcggcaaag gagccgttca
     2761 tttcaataaa ccgggcgacc tcagccatcc cttcctgatt ttccgctttc cagcgttcgg
     2821 cacgcagacg acgggcttca ttctgcatgg ttgtgcttac cgaaccggag atattgacat
     2881 catatatgcc ttgagcaact gatagctgtc gctgtcaact gtcactgtaa tacgctgctt
     2941 catagcatac ctctttttga catacttcgg gtatacatat cagtatatat tcttataccg
     3001 caaaaatcag cgcgcaaata cgcatactgt tatctggctt ttagtaagcc ggatcctcta
     3061 gattacgccc cgcctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga
     3121 catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt
     3181 cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg
     3241 ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat
     3301 tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg
     3361 aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg
     3421 tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct
     3481 caccgtcttt cattgccata cggaattccg gatgagcatt catcaggcgg gcaagaatgt
     3541 gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa
     3601 tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat
     3661 gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca
     3721 ttttagcttc cttagctcct gaaaatctcg acggatccta actcaaaatc cacacattat
     3781 acgagccgga agcataaagt gtaaagcctg ggggtgccta atgcggccgc catagtgact
     3841 ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata
     3901 tattgatatt tatatcattt tacgtttctc gttcaacttt attatacata gttgataatt
     3961 cactggccgt cgttttacgg taccatcgat accgtcgacc tcgacatggt catagctgtt
     4021 gacggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4081 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4141 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4201 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4261 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4321 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4381 agcacgtgtt gaaaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     4441 gaggaactaa accatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4501 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4561 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4621 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4681 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4741 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4801 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4861 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4921 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4981 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5041 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5101 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5161 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5221 ctatcgcctt cttgacgagt tcttctaacg taccggttag taatgagttt aaacggggga
     5281 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     5341 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     5401 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     5461 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     5521 gtcggggcgg caggccctgc catagcagat ccggttaaat caaaagaata gaccgagata
     5581 gggttgagtg ttgttccagt ttggaacaag agtccacgct gtccctgata tctataacaa
     5641 gaaaatatat atataataag ttatcacgta agtagaacat gaaataacaa tataattatc
     5701 gtatgagtta aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt gactcacgcg
     5761 gtcgttatag ttcaaaatca gtgacactta ccgcattgac aagcacgcct cacgggagct
     5821 ccaagcggcg actgagatgt cctaaatgca cagcgacgga ttcgcgctat ttagaaagag
     5881 agagcaatat ttcaagaatg catgcgtcaa ttttacgcag actatctttc tagggttaat
     5941 ctagctgcat caagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     6001 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     6061 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     6121 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     6181 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcag
//
