LOCUS       Exported                6902 bp ds-DNA     circular SYN 19-OCT-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6902)
  AUTHORS   Sohum Mehta
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Nov 8, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6902
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
                     /note="color: #a6acb3"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
                     /note="color: #ffffff"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
                     /note="color: #ffffff"
     CDS             941..2422
                     /codon_start=1
                     /label=iBB-GECO1
                     /note="color: #ff00ff"
                     /note="color: #993366"
                     /translation="MFSVPLTTSAFGHAFLANIEREQIHLAGSIQPHGILLAVKEPDNV
                     VIQASINAAEFLNTNFVVGRPLRDLGGDLPLQILPHLNGPLHLAPMTLRCTVGSPPRRV
                     DCTIHRPSNGGLIVELEPATKATNIAPALVGALHRITSSSSLMGLCDETATIFREITGF
                     DRVMVMRLGALDDLTEEQIAEIKEAFSLFDKDGDGTITTKELGTVFRSLGQNPTEAELQ
                     DLINEVDADGNGIFDFPEFLTMMARKMNDADSEEEIRGAFRVFDKDGNGNIGAAGLRHV
                     MSDLGEKLTDEEVDEMIRVADNDGDGQVNYEEFVQIMTAKGGSVDSSRRKWNKAGHAVR
                     AIGRLSSRRHDLLSECRRADLEAFLGNRYPAYTIPQTARRLYELNRVRLMVDVNYTPVP
                     LEPRISPLNGRDLDMSLSCLRSMSPCHLKYMQDMGVGATLVCSLMVSGRLWGLIVCHHY
                     EPRYVPSHIRAAGEALAEACANRIATLESFAPSQSK"
     CDS             941..1453
                     /codon_start=1
                     /label=iBB-N
                     /note="color: #993366"
                     /note="color: #993366"
                     /translation="MFSVPLTTSAFGHAFLANIEREQIHLAGSIQPHGILLAVKEPDNV
                     VIQASINAAEFLNTNFVVGRPLRDLGGDLPLQILPHLNGPLHLAPMTLRCTVGSPPRRV
                     DCTIHRPSNGGLIVELEPATKATNIAPALVGALHRITSSSSLMGLCDETATIFREITGF
                     DRVMVMRL"
     CDS             1463..1903
                     /codon_start=1
                     /label=CaM
                     /note="color: #942193"
                     /note="color: #993366"
                     /translation="DDLTEEQIAEIKEAFSLFDKDGDGTITTKELGTVFRSLGQNPTEA
                     ELQDLINEVDADGNGIFDFPEFLTMMARKMNDADSEEEIRGAFRVFDKDGNGNIGAAGL
                     RHVMSDLGEKLTDEEVDEMIRVADNDGDGQVNYEEFVQIMTAK"
     CDS             1913..1981
                     /codon_start=1
                     /label=RS20
                     /note="color: #808080"
                     /note="color: #993366"
                     /translation="VDSSRRKWNKAGHAVRAIGRLSS"
     CDS             1994..2422
                     /codon_start=1
                     /label=iBB_C
                     /note="color: #993366"
                     /note="color: #993366"
                     /translation="LLSECRRADLEAFLGNRYPAYTIPQTARRLYELNRVRLMVDVNYT
                     PVPLEPRISPLNGRDLDMSLSCLRSMSPCHLKYMQDMGVGATLVCSLMVSGRLWGLIVC
                     HHYEPRYVPSHIRAAGEALAEACANRIATLESFAPSQSK"
     polyA_signal    2502..2726
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
                     /note="color: #a6acb3"
     rep_origin      2772..3200
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
                     /note="color: #ffff00"
     promoter        3214..3543
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
                     /note="color: #ffffff"
     rep_origin      3394..3529
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
                     /note="color: #ffff00"
     CDS             3610..4404
                     /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))"
                     /note="color: #ccffcc"
                     /note="color: #993366"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4578..4699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
                     /note="color: #a6acb3"
     primer_bind     complement(4748..4764)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    4772..4788
                     /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)."
                     /note="color: #31849b"
                     /note="color: #31849b"
     promoter        complement(4796..4826)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                     1: 4796 .. 4802 / #ffffff / -10
                     2: 4803 .. 4820 / #ffffff
                     3: 4821 .. 4826 / #ffffff / -35"
                     /note="color: #ffffff; direction: LEFT"
     protein_bind    4841..4862
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /note="color: #31849b"
                     /note="color: #31849b"
     rep_origin      complement(5150..5735)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
                     /note="color: #ffff00"
     CDS             complement(5906..6766)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                     1: 5906 .. 6697 / #ccffcc
                     2: 6698 .. 6766 / #ccffcc / signal sequence
                     Cleavage site after base 6697"
                     /note="color: #993366"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6767..6871)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgttctcgg taccgctgac
      961 tacctcagct ttcggccacg cgtttctggc taacattgaa cgtgagcaga tccacctggc
     1021 gggctccatt cagccgcacg gtatcctgct ggctgttaaa gagcctgaca acgtggtgat
     1081 ccaggcttct attaacgctg cggagttcct gaacaccaac tttgttgttg gccgtccgct
     1141 gcgtgacctg ggcggcgatc tgcctttgca gatcctgccg cacctgaacg gcccgctgca
     1201 cctggctccg atgaccctgc gttgtactgt gggttctccg ccgcgtcgtg tggactgtac
     1261 cattcatcga ccgtctaacg gcggcctgat cgtagaactg gaaccagcaa ccaaggccac
     1321 taacattgcg ccggctctgg tcggtgcgct tcatcgtatc acttcttcat cctccctgat
     1381 gggcctgtgt gacgaaaccg cgactatttt ccgtgagatc actggtttcg accgtgtgat
     1441 ggtaatgcgt ctcggcgcgc ttgacgatct gactgaagag cagatcgcag agattaaaga
     1501 ggctttctcc ctatttgaca aggacgggga cgggacgata acaaccaagg agctggggac
     1561 ggtgttccgg tctctggggc agaaccccac agaagcagag ctgcaggact taatcaacga
     1621 ggtagatgcc gatggcaacg gcatattcga cttccctgag ttcctgacga tgatggcaag
     1681 gaaaatgaat gacgcagaca gtgaagagga aattagagga gcgttccgcg tgtttgataa
     1741 ggacggcaat ggcaacatcg gcgcagcagg gcttcgccac gtgatgtcag accttggtga
     1801 gaagttaact gatgaggagg ttgatgaaat gatcagggta gcggacaacg atggggatgg
     1861 tcaggtaaac tacgaagagt ttgtacaaat catgacagcg aagggaggtt ctgtagattc
     1921 atcacgtcgt aagtggaata aggcaggaca cgcagtcaga gctataggtc ggctgagctc
     1981 gcgtaggcat gatttgcttt ccgaatgtcg tcgtgcggac ctggaagcgt tcctgggtaa
     2041 ccgctacccg gcgtatacta ttccgcagac cgctcgtcgc ctgtacgaac tcaaccgtgt
     2101 tcgcctgatg gtagatgtga actatactcc ggttcctcta gagccgcgca tcagcccgct
     2161 gaacggtcgt gatctggata tgtccctgtc ttgcctgcgc tctatgtccc cgtgccacct
     2221 gaaatacatg caggacatgg gcgtgggcgc aaccctggtt tgctctctga tggtgtctgg
     2281 tcgtctgtgg ggtctgatcg tttgccacca ctacgaaccg cgctacgttc cgtcccacat
     2341 tcgcgctgct ggcgaagcgc tggcggaagc atgtgcgaac cgcatcgcga cgctggagag
     2401 ctttgcaccg tctcagtcca aatgagaatt ctgcagatat ccagcacagt ggcggccgct
     2461 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     2521 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2581 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2641 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     2701 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     2761 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2821 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2881 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2941 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3001 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3061 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3121 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3181 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3241 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3301 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3361 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3421 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3481 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3541 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     3601 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3661 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3721 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3781 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3841 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3901 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3961 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4021 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4081 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4141 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4201 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4261 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4321 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4381 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4441 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4501 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4561 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4621 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4681 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4741 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4801 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4861 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4921 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4981 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5041 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5101 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5161 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5221 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5281 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5341 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5401 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5461 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5521 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5581 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5641 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     5701 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5761 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5821 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5881 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5941 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6001 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6061 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6121 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6181 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6241 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6301 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6361 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6421 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6481 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6541 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6601 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6661 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6721 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6781 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6841 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6901 tc
//
